Stem cell-based bone and dental regeneration: a view of microenvironmental modulation

In modern medicine,bone and dental loss and defects are common and widespread morbidities,for which regenerative therapy has shown great promise.Mesenchymal stem cells,obtained from various sources and playing an essential role in organ development and postnatal repair,have exhibited enormous potential for regenerating bone and dental tissue.Currently,mesenchymal stem cells (MSCs)-based bone and dental regeneration mainly includes two strategies: the rescue or mobilization of endogenous MSCs and the application of exogenous MSCs in cytotherapy or tissue engineering.Nevertheless,the efficacy of MSCbased regeneration is not always fulfilled,especially in diseased microenvironments.Specifically,the diseased microenvironment not only impairs the regenerative potential of resident MSCs but also controls the therapeutic efficacy of exogenous MSCs,both as donors and recipients.Accordingly,approaches targeting a diseased microenvironment have been established,including improving the diseased niche to restore endogenous MSCs,enhancing MSC resistance to a diseased microenvironment and renormalizing the microenvironment to guarantee MSC-mediated therapies.Moreover,the application of extracellular vesicles (EVs) as cell-free therapy has emerged as a promising therapeutic strategy.In this review,we summarize current knowledge regarding the tactics of MSC-based bone and dental regeneration and the decisive role of the microenvironment,emphasizing the therapeutic potential of microenvironment-targeting strategies in bone and dental regenerative medicine.

Anti-proteolytic capacity and bonding durability of proanthocyanidin-biomodified demineralized dentin matrix

Our previous studies showed that biomodification of demineralized dentin collagen with proanthocyanidin（PA） for a clinically practical duration improves the mechanical properties of the dentin matrix and the immediate resin–dentin bond strength. The present study sought to evaluate the ability of PA biomodification to reduce collagenase-induced biodegradation of demineralized dentin matrix and dentin/adhesive interfaces in a clinically relevant manner. The effects of collagenolytic and gelatinolytic activity on PA-biomodified demineralized dentin matrix were analysed by hydroxyproline assay and gelatin zymography. Then, resin-/dentin-bonded specimens were prepared and challenged with bacterial collagenases. Dentin treated with 2% chlorhexidine and untreated dentin were used as a positive and negative control, respectively. Collagen biodegradation, the microtensile bond strengths of bonded specimens and the micromorphologies of the fractured interfaces were assessed. The results revealed that both collagenolytic and gelatinolytic activity on demineralized dentin were notably inhibited in the PA-biomodified groups, irrespective of PA concentration and biomodification duration. When challenged with exogenous collagenases, PA-biomodified bonded specimens exhibited significantly less biodegradation and maintained higher bond strengths than the untreated control. These results suggest that PA biomodification was effective at inhibiting proteolytic activity on demineralized dentin matrix and at stabilizing the adhesive/dentin interface against enzymatic degradation, is a new concept that has the potential to improve bonding durability.

Development and application of rapid rehabilitation system for reconstruction of maxillofacial soft-tissue defects related to war and traumatic injuries

Background: Maxillofacial war injuries usually cause severe facial organ defects and deformities, handicapping the patient's daily activities, even result in a tendency to commit suicide. The application of maxillofacial prosthesis is an alternative to surgery in functional–aesthetic facial reconstruction. Computer aided design and computer aided manufacturing has opened up a new approach to the fabrication of maxillofacial prosthesis. An intelligentized rapid simulative design and manufacture system for prosthesis was developed to facilitate the prosthesis fabrication procedure.Methods: Maxillofacial prosthesis rapid simulation design and rapid fabrication system consists of three components: digital impression, intelligentized prosthesis designing, and rapid manufacturing. The patients' maxillofacial digital impressions were taken with Structured-light 3D scanner; and then the 3D model of prostheses and their negative molds could be designed in specific software; finally, with the resin molds fabricated by rapid prototyping machine, the prostheses could be produced directly and quickly.Results: Fifteen patients of maxillofacial defect caused by traumatic injuries received prosthesis rehabilitation provided by the established system. The contour of the prostheses coordinated properly with the appearance of the patients, and the uniform-thickness border sealed well to adjacent tissues. All the patients were satisfied with their prostheses.Conclusions: The rapid simulative rehabilitation system of maxillofacial defects has been approaching completion. It could provide advanced technological reservation for the Army in the issue of maxillofacial defect rehabilitation.

Visualization analysis of research frontiers and trends in nerve regeneration and osseoperception in the repair of tooth loss

This study analyzed 85 articles indexed by the Web of Science concerning nerve regeneration and osseoperception during tooth loss repair. Using the Web of Science database and Citespace Ⅲ software, a document co-citation network map was drawn by document co-citation analysis and word frequency analysis methods. Combined with emergent node secondary literature retrieval, subject headings with apparent changing word frequency trends were retrieved so as to identify research frontiers and development trends. Research frontiers and hotspots for neuronal calcium sensor protein were quantitatively explored to forecast future research developments in nerve regeneration and osseoperception during repair of tooth loss.

Identification of ITGA3 as an Oncogene in Human Tongue Cancer via Integrated Bioinformatics Analysis

Human tongue cancer （TC） is an aggressive malignancy with a very poor prognosis. There is an urgent need to elucidate the underlying molecular mechanisms involved in TC progression, mRNA expression profiles play a vital role in the exploration of cancer-related genes. Therefore, the purpose of our study was to identify the progression associated candidate genes of TC by bioinformatics analysis. Five microarray datasets of TC samples were downloaded from the Gene Expression Onmibus （GEO） database and the data of 133 TC patients were screened from The Cancer Genome Atlas （TCGA） head and neck squamous cell carcinoma （HNSC） database. The integrated analysis of five microarray datasets and the RNA sequencing data of TC samples in TCGA-HNSC was performed to obtain 1023 overlapping differentially expressed genes （DEGs） in TC and adjacent normal tissue （ANT） samples. Next, Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis was conducted to enrich the significant pathways of the 1023 DEGs and PI3K- Akt signaling pathway （P=0.011） was selected to be the candidate pathway. A total of 23 DEGs with ｜log2 fold change （FC）｜ ≥1.0 in phosphatidylinositol 3-kinase-serine/threonine kinase （PI3K-Akt） signaling pathway were subjected to survival analysis of 125 eligible TC samples in TCGA database, indicating increased integrin-α3 gene （ITGA3） expression was significantly associated with poorer prognosis. Taken together, our study suggested ITGA3 may facilitate the development of TC via activating PI3K-Akt signaling pathway.

Targeted inhibition of osteoclastogenesis reveals the pathogenesis and therapeutics of bone loss under sympathetic neurostress

Sympathetic cues via the adrenergic signaling critically regulate bone homeostasis and contribute to neurostress-induced bone loss,but the mechanisms and therapeutics remain incompletely elucidated.Here,we reveal an osteoclastogenesis-centered functionally important osteopenic pathogenesis under sympatho-adrenergic activation with characterized micro RNA response and efficient therapeutics.We discovered that osteoclastic mi R-21 was tightly regulated by sympatho-adrenergic cues downstream theβ2-adrenergic receptor(β2AR)signaling,critically modulated osteoclastogenesis in vivo by inhibiting programmed cell death 4(Pdcd4),and mediated detrimental effects of both isoproterenol(ISO)and chronic variable stress(CVS)on bone.Intriguingly,without affecting osteoblastic bone formation,bone protection against ISO and CVS was sufficiently achieved by a(D-Asp8)-lipid nanoparticle-mediated targeted inhibition of osteoclastic mi R-21 or by clinically relevant drugs to suppress osteoclastogenesis.Collectively,these results unravel a previously underdetermined molecular and functional paradigm that osteoclastogenesis crucially contributes to sympatho-adrenergic regulation of bone and establish multiple targeted therapeutic strategies to counteract osteopenias under stresses.

A 3D-printed molybdenum-containing scaffold exerts dual pro-osteogenic and anti-osteoclastogenic effects to facilitate alveolar bone repair

The positive regulation of bone-forming osteoblast activity and the negative feedback regulation of osteoclastic activity are equally important in strategies to achieve successful alveolar bone regeneration. Here, a molybdenum(Mo)-containing bioactive glass ceramic scaffold with solid-strut-packed structures(Mo-scaffold) was printed, and its ability to regulate pro-osteogenic and antiosteoclastogenic cellular responses was evaluated in vitro and in vivo. We found that extracts derived from Mo-scaffold(Moextracts) strongly stimulated osteogenic differentiation of bone marrow mesenchymal stem cells and inhibited differentiation of osteoclast progenitors. The identified comodulatory effect was further demonstrated to arise from Mo ions in the Mo-extract,wherein Mo ions suppressed osteoclastic differentiation by scavenging reactive oxygen species(ROS) and inhibiting mitochondrial biogenesis in osteoclasts. Consistent with the in vitro findings, the Mo-scaffold was found to significantly promote osteoblastmediated bone formation and inhibit osteoclast-mediated bone resorption throughout the bone healing process, leading to enhanced bone regeneration. In combination with our previous finding that Mo ions participate in material-mediated immunomodulation, this study offers the new insight that Mo ions facilitate bone repair by comodulating the balance between bone formation and resorption. Our findings suggest that Mo ions are multifunctional cellular modulators that can potentially be used in biomaterial design and bone tissue engineering.

Expert consensus on early childhood caries management

Early childhood caries(ECC)is a significant chronic disease of childhood and a rising public health burden worldwide.ECC may cause a higher risk of new caries lesions in both primary and permanent dentition,affecting lifelong oral health.The occurrence of ECC has been closely related to the core microbiome change in the oral cavity,which may be influenced by diet habits,oral health management,fluoride use,and dental manipulations.So,it is essential to improve parental oral health and awareness of health care,to establish a dental home at the early stage of childhood,and make an individualized caries management plan.Dental interventions according to the minimally invasive concept should be carried out to treat dental caries.This expert consensus mainly discusses the etiology of ECC,caries-risk assessment of children,prevention and treatment plan of ECC,aiming to achieve lifelong oral health.

Third molar-related knowledge,attitudes,behaviors,and medical history of 904 Chinese adults:a cross-sectional survey

This study investigated the perceptions and medical history of third molars(M3s)and assessed the prevalence of visible M3s(V-M3s)among 904 Chinese adults.The enrolled participants were interviewed to complete a structural questionnaire focused on sociodemographic information and their understanding of,attitudes toward,behaviors regarding,and medical history with respect to M3s.In addition,the number of V-M3s in the cohort was determined by oral examination.Logistic regression analysis was performed to explore the association between individuals'sociodemographic characteristics and their perception of M3s or the presence of V-M3s.The Chi-square test was used to compare the actions taken against symptomatic M3s and the corresponding outcomes among different groups divided according to respondents'sociodemographic factors.In total,904 completed questionnaires were gathered and analyzed.Nearly half(43.9%)of the respondents knew nothing about M3s,and only 12.7%provided correct answers to all the questions asked.Male sex,older age,occupation involving physical labor,and no previous dental experience were active factors in unawareness of M3s.Male sex was also significantly associated with the presence of at least one V-M3 and negative behavior about symptomatic M3s.In terms of medical history,192 participants reported having had at least one M3 extracted(438 in total),and 72.6%of the M3s were removed due to the presence of related symptoms or pathologies.In conclusion,the population investigated had a shortage of knowledge about M3s and adopted negative attitudes and actions about M3-related problems.

Surface demineralized freeze-dried bone allograft followed by reimplantation in a failed mandibular dental implant

The removal of a failed implant with high torque causes significant damage to the surrounding tissue,compromising bone regeneration and subsequent osseointegration in the defect area.Here,we report a case of carrier screw fracture followed by immediate implant removal,bone grafting and delayed reimplantation.A dental implant with a fractured central carrier screw was removed using the bur-forceps technique.The resulting three-wall bone defect was filled with granular surface demineralized freeze-dried bone allograft(SD-FDBA).Cone-beam computerized tomography was performed at 1week,6months and 15months postoperatively and standardized for quantitative evaluation.The alveolar bone width and height at 15months post-surgery were about 91%of the original values,with a slightly lower bone density,calculated using the gray value ratio.The graft site was reopened and was found to be completely healed with dense and vascularized bone along with some residual bone graft.Reimplantation followed by restoration was performed 8months later.The quality of regenerated bone following SD-FDBA grafting was adequate for osseointegration and longterm implant success.The excellent osteogenic properties of SD-FDBA are attributed to its human origin,cortical bone-like structure,partly demineralized surfaces and bone morphogenetic protein-2-containing nature.Further investigation with more cases and longer follow-up was required to confirm the final clinical effect.

Self-adaptive hydrogel for breast cancer therapy via accurate tumor elimination and on-demand adipose tissue regeneration

The irregular defects and residual tumor tissue after surgery are challenges for effective breast cancer treatment.Herein,a smart hydrogel with self-adaptable size and dual responsive cargos release was fabricated to treat breast cancer via accurate tumor elimination,on-demand adipose tissue regeneration and effective infection inhibition.The hydrogel consisted of thiol groups ended polyethylene glycol(SH-PEG-SH)and doxorubicin encapsulated mesoporous silica nanocarriers(DOX@MSNs)double crosslinked hyaluronic acid(HA)after loading of antibacterial peptides(AP)and adipose-derived stem cells(ADSCs).A pH-cleavable unsaturated amide bond was pre-introduced between MSNs and HA frame to perform the tumor-specific acidic environment dependent DOX@MSNs release,meanwhile an esterase degradable glyceryl dimethacrylate cap was grafted on MSNs,which contributed to the selective chemotherapy in tumor cells with over-expressed esterase.The bond cleavage between MSNs and HA would also cause the swelling of the hydrogel,which not only provide sufficient space for the growth of ADSCs,but allows the hydrogel to fully fill the irregular defects generated by surgery and residual tumor atrophy,resulting in the on-demand regeneration of adipose tissue.Moreover,the sustained release of AP could be simultaneously triggered along with the size change of hydrogel,which further avoided bacterial infection to promote tissue regeneration.

A retrospective study to compare the treatment outcomes with and without surgical navigation for fracture of the orbital wall

Purpose To evaluate the outcomes with and without aid of a computer-assisted surgical navigation system(CASNS)for treatment of unilateral orbital wall fracture(OWF).Methods Patients who came to our hospital for repairing unilateral traumatic OWF from 2014 to 2017 were included in this study.The patients were divided into the navigation group who accepted orbital wall reconstruction aided by CASNS and the conventional group.We evaluated the surgical precision in the navigation group by analyzing the difference between actual postoperative computed tomography data and preoperative virtual surgical plan through color order ratios.We also compared the duration of surgery,enophthalmos correction,restoration of orbital volumes,and improvement of clinical symptoms in both groups systemically.Quantitative data were presented as mean±SD.Significance was determined by the two-sample t-test using SPSS Version 19.0 A p<0.05 was considered statistically significant.Results Seventy patients with unilateral OWF were included in the study cohort.The mean difference between preoperative virtual planning and actual reconstruction outcome was(0.869±0.472)mm,which means the reconstruction result could match the navigation planning accurately.The mean duration of surgery in the navigation group was shorter than it is in the control group,but not significantly.Discrepancies between the reconstructed and unaffected orbital-cavity volume and eyeball projection in the navigation group were significantly less than that in the conventional group.One patient had remnant diplopia and two patients had enophthalmos after surgery in the navigation group;two patients had postoperative diplopia and four patients had postoperative enophthalmos in the conventional group.Conclusion Compare with the conventional treatment for OWF,the use of CASNS can provide a significantly better surgical precision,greater improvements in orbital-cavity volume and eyeball projection,and better clinical results,without increasing the duration of surgery.

Microstructure and Mechanical Properties of Al_2O_3/ZrO_2 Directionally Solidified Eutectic Ceramic Prepared by Laser 3D Printing

Directionally solidified eutectic ceramics such as Al2O3/ZrO2 are promising structural materials for ap- plications in harsh environment with an ultrahigh temperature. In this work, through adopting assistant heating laser 3D printing, Al2O3/ZrO2 eutectic samples were manufactured with suppressing the forma- tion of cracks. The dependence of the average rod spacing （λav） on the scanning rate （V） follows a relation with λavV0.5 = 1 μm1.5 s^-0.5. Typical eutectic microstructures, so-called complex regular, were analyzed with respect to its evolution with modulating the growth conditions. Formation mechanism of the solidifi- cation defect, shrinkage porosity, was discussed and the defect is found to be significantly suppressed by optimizing the solidification parameters. The maximum hardness and fracture toughness are mea- sured to he 16.7 GPa and 4.5 MPa m1/2, respectively. The interplay between the propagation of cracks and the Al2O3/ZrO2 interface is discussed.

The influence, of nanotopography on organelle organization and communication

Cellular differentiation can be affected by the extracellular environment, particularly extracellular substrates. The nanotopography of the substrate may be involved in the mechanisms of cellular differentiation in vivo. Organelles are major players in various cellular functions; however, the influence of nano- topography on organelles has not yet been elucidated. In the present study, a micropit-nanotube topography （MNT） was fabricated on the titanium surface, and organelle-specific fluorescent probes were used to detect the intracellular organelle organization of MG63 cells. Communication between organelles, identified by organelle-specific GTPase expression, was evaluated by quantitative polymerase chain reaction and western blotting. Transmission electron microscopy was performed to evaluate the organelle structure. There were no significant differences in organelle distribution or number between the MNT and flat surface. However, organelle-specific GTPases on the MNT were dramatically downregulated. In addition, obvious endoplasmic reticulum lumen dilation was observed on the MNT surface, and the unfolded protein response （UPR） was also initiated. Regarding the relationships among organelle trafficking, UPR, and osteogenic differentiation, our findings may provide important insights into the signal transduction induced by nanotopography.

Immobilization of heparin on decellularized kidney scaffold to construct microenvironment for antithrombosis and inducing reendothelialization

In recent years, rapid development of tissue engineering technology provides possibilities for the construction of artificial tissues or organs. In construction of engineered kidneys, researchers used native decellularized extracellular matrix(ECM) as the scaffolds to recellularization. However, thrombosis has been a great issue that hinders the progress of transplantation in vivo. In this study, heparin was immobilized to the collagen part of decellularized scaffold with collagen-binding peptide(CBP). Through the anticoagulant and endothelial cell reperfusion experiments, it can be demonstrated that the heparinized scaffolds absorbed less platelets and red blood cells which can effectively reduce the formation of thrombosis. Moreover, it is conducive to longterm adhesion of endothelial cells which is important for the formation of subsequent vascularization. Taken together, our results reveal that the whole kidney can be modified by CBP-heparin composite to reduce the thrombosis and provide the better conditions for neovascularization.

Comparison of Different Periodontal Healing of Critical Size Noncontained and Contained Intrabony Defects in Beagles

Background： Regenerative techniques help promote the formation of new attachment and bone filling in periodontal detects. However, the dimensions of intraosseous defects are a key determinant of periodontal regeneration outcomes. In this study, we evaluated the efficacy of use of anorganic bovine bone （ABB） graft in combination with collagen membrane （CM）, to facilitate healing ofnoncontained （1-wall） and contained （3-wall） critical size periodontal defects. Methods： The study began on March 2013, and was completed on May 2014. One-wall （7 mm × 4 mm） and 3-wall （5 mm × 4 mm） intrabony periodontal defects were surgically created bilaterally in the mandibular third premolars and first molars in eight beagles. The defects were treated with ABB in combination with CM （ABB ＋ CM group） or open flap debridement （OFD group）. The animals were euthanized at 8-week postsurgery for histological analysis. Two independent Student＇s t-tests （1-wall [ABB ＋ CM] vs. 1-wall [OFD] and 3-wall [ABB ＋ CM] vs. 3-wall [OFD]） were used to assess between-group differences. Results： The mean new bone height in both 1- and 3-wall intrabony defects in the ABB ＋ CM group was significantly greater than that in the OFD group （1-wall： 4.99± 0.70 mm vs. 3.01 ± 0.37 mm, P 〈 0.05; 3-wall： 3.11±0.59 mm vs. 2.08± 0.24 mm, P 〈 0.05）. The mean new cementum in 1-wall intrabony defects in the ABB ＋ CM group was significantly greater than that in their counterparts in the OFD group （5.08 ±0.68 mm vs. 1.16 ± 0.38 mm; P 〈 0.05）. Likewise, only the 1-wall intrabony defect model showed a significant difference with respect to junctional epithelium between ABB ＋ CM and OFD groups （0.67 ±0.23 mm vs. 1.12 ± 0.28 mm, P 〈 0.05）. Conclusions： One-wall intrabony defects treated with ABB and CM did not show less periodontal regeneration than that in 3-wall intrabony defect. The noncontained l-wall intrabony defect might be a more discriminative defect model for further research into periodontal regeneration.

In vivo biocompatibility and degradability of a Zn-Mg-Fe alloy osteosynthesis system

Zinc is generally considered to be one of the most promising materials to be used in biodegradable implants,and many zinc alloys have been optimized to improve implant biocompatibility,degradation,and mechanical properties.However,long-term degradation leads to the prolonged presence of degradation products,which risks foreign body reactions.Herein,we investigated the in vivo biocompatibility and degradation of a biodegradable Zn-Mg-Fe alloy osteosynthesis system in the frontal bone,mandible,and femur in beagles for 1 year.Results of the routine blood,biochemical,trace element,and histological analyses of multiple organs,peripheral blood CD4/CD8a levels,and serum interleukin 2 and 4 levels showed good biocompatibility of the Zn-Mg-Fe alloy.Zinc content analysis revealed zinc accumulation in adjacent bone tissue,but not in the liver,kidney,and spleen,which was related to the degradation of the Zn-Mg-Fe alloy.The alloy demonstrated a uniform slowing degradation rate in vivo.No degradation differences in the frontal bone,mandible,and femur were observed.The degradation products included zinc oxide[ZnO],zinc hydroxide[Zn(OH)_(2)],hydrozincite[Zn_(5)(OH)_(6)(CO_(3))_(2)],and hopeite[Zn_(3)(PO_(4))_(2)⋅4H_(2)O].The good biocompatibility and degradation properties of the Zn-Mg-Fe alloy render it a very attractive osteosynthesis system for clinical applications.

Porous bio-high entropy alloy scaffolds fabricated by direct ink writing

Porous tantalum-titanium-niobium-zirconium(Ta-Ti-Nb-Zr)bio-high entropy alloy(bioHEA)scaffolds are fabricated using direct ink writing 3D printing technology in this study.A composite ink is prepared using four metal powders as raw materials:Ta,Ti,Nb and Zr.Ink extrusion is used to build 3D scaf-folds with interconnected porous structures at room temperature,which are then sintered in a vacuum environment.The interdiffusion of metal elements yields porous bioHEA scaffolds with a body-centered cubic(BCC)structure.The fabricated scaffolds have uniform compositions with a significant alloying ef-fect and good biocompatibility.The scaffolds have a compressive strength of 70.08-149.95 MPa and an elastic modulus of 0.18-0.64 GPa,indicating that the mechanical properties can be controlled over a wide range.The scaffolds have a compressive strength close to that of human cortical bone and thus meet the requirements for porous structure characteristics and biological and mechanical properties of orthopedic implants.

Dental stem cell and dental tissue regeneration

The teeth are highly differentiated chewing organs formed by the development of tooth germ tissue located in the jaw and consist of the enamel, dentin, cementum, pulp, and periodontal tissue. Moreover, the teeth have a complicated regulatory mechanism, special histologic origin, diverse structure, and important function in mastication, articulation, and aesthetics. These characteristics, to a certain extent, greatly complicate the research in tooth regeneration. Recently, new ideas for tooth and tissue regeneration have begun to appear with rapid developments in the theories and technologies in tissue engineering. Numerous types of stem cells have been isolated from dental tissue, such as dental pulp stem cells (DPSCs), stem cells isolated from human pulp of exfoliated deciduous teeth (SHED), periodontal ligament stem cells (PDLSCs), stem cells from apical papilla (SCAPs), and dental follicle cells (DFCs). All these cells can regenerate the tissue of tooth. This review outlines the cell types and strategies of stem cell therapy applied in tooth regeneration, in order to provide theoretical basis for clinical treatments.

Epithelial defect repair in the auricle and auditory meatus by grafting with cultured adipose-derived mesenchymal stem cell aggregate-extracellular matrix

Background: Several patients experience persistent otorrhea after a flawless surgical procedure because of insufficient epithelial healing. Several efforts, such as autologous tissue allograft and xenograft, have been made to halt otorrhea. However, a stable technology to induce temporal epithelial repair is yet to be established. Therefore, this study aims to investigate whether implantation of seeding adipose-derived mesenchymal stem cell (ADMSC) aggregates on extracellular matrix (ECM;herein, ADMSC aggregate-ECM) into damaged skin wound promotes skin regeneration. Methods: ADMSC aggregate-ECM was prepared using a previously described procedure that isolated ADMSCs from rabbits and applied to the auricle and auditory meatus wound beds of New Zealand white rabbits. Wound healing was assessed by general observation and hematoxylin and eosin (H&E) staining. Secretion of growth factor of the tissue was evaluated by western blotting. Two other groups, namely, ECM and control, were used. Comparisons of three groups were conducted by one-way analysis of variance analysis. Results: ADMSCs adhered tightly to the ECM and quickly formed cell sheets. At 2 weeks, general observation and H&E staining indicated that the wound healing rates in the ADMSC aggregate-ECM (69.02±6.36%) and ECM (59.32 + 4.10%) groups were higher than that in the control group (43.74± 12.15%;P=0.005, P<0.001, respectively) in ear auricle excisional wounds. At 7 weeks, The scar elevation index was evidently reduced in the ADMSC aggregate-ECM (2.08±0.87) and ECM (2.31 ±0.33) groups compared with the control group (4.06 ±0.45;P < 0.001, P < 0.001, respectively). In addition, the scar elevation index of the ADMSC aggregate-ECM group reached the lowest rate 4 weeks in advance. In auditory meatus excisional wounds, the ADMSC aggregate-ECM group had the largest range of normal skin-like structure at 4 weeks. The ADMSC aggregate-ECM and ECM groups secreted increased amounts of growth factors that contributed to skin regeneration at weeks 1 and 2, respectively. Conclusions: ADMSC aggregate-ECM and ECM are effective repair materials for wound healing, especially ADMSC aggregate- ECM. This approach will provide a meaningful experimental basis for mastoid epithelium repair in subsequent clinical trials.