Surface-modified Biphasic Calcium Phosphate/Poly (L-lactide) Biocomposite

Biphasic calcium phosphate （BCP） powders were prepared by hydrolyzation proc-ess and surface-modified by directly grafted L-lactide （LLA） onto the surface of BCP through a chemical linkage. The grafting ratio of organic groups was 9 wt%. After surface modification, the surface of BCP powders was covered by the lamella-shaped crystal. Poly （L-lactide） was mixed with BCP to form the BCP/PLLA biocomposite. Modified BCP （mBCP） particles could be uniformly dis-persed in PLLA matrix. The compressive strength of the mBCP/PLLA composite is 115 MPa, 28% higher than that of unmodified-BCP/PLLA composite. The improved mechanical strength is attributed to the enhanced adhesion between the inorganic BCP filler and the organic PLLA matrix.

Graphene oxide coated three-dimensional printed biphasic calcium phosphate scaffold for angiogenic and osteogenic synergy in repairing critical-size bone defect

The custom-tailored medicine requires a developmental strategy that integrates excellent osteogene-sis with mechanical stability to enhance the reconstruction of the critical-size bone defect(CSBD)and the healing process in weight-bearing bone.We prepared three-dimensional(3D)printed biphasic cal-cium phosphate(BCP)scaffolds composited with nano-graphene oxide(GO).The biological effects of the GO/BCP composite scaffolds could induce the differentiation of rat bone marrow stem cells(BM-SCs)and the migration of human umbilical vein endothelial cells(HUVECs)for bone repair.The proper ratio of GO in the composite scaffold regulated the composites’surface roughness and hydrophilicity to a suitable range for the adhesion and proliferation of BMSCs and HUVECs.Besides,the GO/BCP composite scaffold increased osteogenesis and angiogenesis by activating BMP-2,RUNX-2,Smad1/4,and VEGF.The customized intramedullary nail combined with GO/BCP scaffold was applied to repair CSBD(2.0 cm in length)in a beagle femur model.This fixation strategy was confirmed by finite element analysis.In vivo,the results indicated that the custom-made internal fixation provided sufficient stability in the early stage,ensuring bone healing in a considerable mechanical environment.At 9 months postoperatively,longitudi-nal bony union and blood vessels in osteon were observed in the CSBD area with partial degradation in the 0.3%GO/BCP group.In the three-point bending test,the ultimate load of 0.3%GO/BCP group reached over 50%of the normal femur at 9 months after repair.These results showed a promising application of osteogenic GO/BCP scaffold and custom-made intramedullary nails in repairing CSBD of the beagle femur.This effective strategy could provide an option to treat the clinical CSBD in weight-bearing bones.

Degradation and biological performance of porous osteomimetic biphasic calcium phosphate in vitro and in vivo

The combination between biphasic calcium phosphate(BCP)and the osteomimetic porous microstructure obtained via freeze casting is hoped to achieve excellent bone regeneration,while the effects of HA and b-TCP ratio changes on the degradation and biological performance of the BCP scaffolds with this unique microstructure need to be determined.Here,we prepared the osteomimetic BCP scaffolds with different HA/b-TCP ratios(HA30/b-TCP70,HA50/b-TCP50,HA70/b-TCP30)and the effects of different HA/b-TCPHA/b-TCP ratios on the degradation and biological performance were studied in vitro and vivo.These BCP scaffolds with different HA/b-TCP ratios exhibited similar microstructure,mechanical performance,and protein absorption capability,while HA70/b-TCP30 BCP scaffolds showed an advisable degradation rate.Study in vitro confirmed the bio-compatibility and promotion on the proliferation,differentiation of MG63 cells in the porous osteomimetic BCP scaffolds with a HA/b-TCP ratio at 30:70.Implantation experiments also showed that the porous osteomimetic BCP scaffolds with a HA/b-TCP ratio at 30:70 had excellent bone regeneration capacity and proper degradation rate compatible with bone growth.These results reveal that the porous osteomimetic BCP scaffold with a HA/b-TCP ratio at 30:70 is a potential candidate of biodegradable bone substitutes used for bone repair.

Biphasic calcium phosphate nano-composite scaffolds reinforced with bioglass provide a synthetic alternative to autografts in a canine tibiofibula defect model

Background Bone grafting is commonly used to repair bone defects.As the porosity of the graft scaffold increases,bone formation increases,but the strength decreases.Early attempts to engineer materials were not able to resolve this problem.In recent years,nanomaterials have demonstrated the unique ability to improve the material strength and toughness while stimulating new bone formation.In our previous studies,we synthesized a nano-scale material by reinforcing a porous β-tricalcium phosphate （β-TCP） ceramic scaffold with Na2O-MgO-P2O5-CaO bioglass （β-TCP/BG）.However,the in vivo effects of the β-TCP/BG scaffold on bone repair remain unknown.Methods We investigated the efficacy of β-TCP/BG scaffolds compared to autografts in a canine tibioflbula defect model.The tibioflbula defects were created in the right legs of 12 dogs,which were randomly assigned to either the scaffold group or the autograft group （six dogs per group）.Radiographic evaluation was performed at 0,4,8,and 12 weeks post-surgery.The involved tibias were extracted at 12 weeks and were tested to failure via a three-point bending.After the biomechanical analysis,specimens were subsequently processed for scanning electron microscopy analysis and histological evaluations.Results Radiographic evaluation at 12 weeks post-operation revealed many newly formed osseous calluses and bony unions in both groups.Both the maximum force and break force in the scaffold group （n=6） were comparable to those in the autograft group （n=6,P ＞0.05）,suggesting that the tissue-engineered bone repair achieved similar biomechanical properties to autograft bone repair.At 12 weeks post-operation,obvious new bone and blood vessel formations were observed in the artificial bone of the experimental group.Conclusions The results demonstrated that new bone formation and high bone strength were achieved in the β-TCP/ BG scaffold group,and suggested that the β-TCP/BG scaffold could be used as a synthetic alternative to autografts for the repair of bone defects.

Whisker of biphasic calcium phosphate ceramics: Osteoimmunomodulatory behaviors

Immune systems play a critical role in the regulation of bone formation and homeostasis,which arouses a growing interest in the development of biomaterials that can modulate both immune response and osteogenesis.In this study,biphasic calcium phosphate(BCP)ceramics were modified with different whiskered surface,and their effects on macrophage polarization and functional status were investigated.The results showed that compared to BCP-W ceramics with long and solid whiskers,BCPHW ceramics with short and hollow whiskers surface were conducive to protein adsorption and macrophage elongation.Furthermore,BCP-HW ceramics down-regulated the expression of M1 macrophage markers(Il1β,Tnfα,and iNos),promoted the expression of M2 macrophage markers(Il10 and Arg)and growth factors(Tgfβ1 and Bmp2),which might be attributed to the differential integrin expression regulated by different whisker structures.The conditioned medium derived from the supernatant of macrophage/whiskered ceramic co-culture was further used to culture MC3T3-E1 pre-osteoblasts to evaluate the effects of whiskered ceramic-mediated macrophage secretion on osteogenesis in vitro.Compared with BCP-W ones,the secretion pattern induced by BCP-HW ceramics could promote the expression of bone markers in pre-osteoblasts,which might due to the activation of intracellular signaling cascades like BMP/Smad and TGF-β/Smad signaling pathways.A murine intramuscular implantation model suggested that after implantation for 1,2,and 3 weeks,BCP-HW ceramics drove the switch of macrophages to ARG+wound-healing M2 phenotype,while BCP-W ceramics increased the proportion of iNOS+M1 inflammatory macrophages.At 2 months,only BCP-HW could induce ectopic bone formation.Taken together,these results indicated that BCP ceramics with hollow whiskers were capable of creating a proper inflammatory microenvironment to induce bone formation.These whiskered BCP ceramics with good osteo-immunomodulatory capacity hold promise in serving as bone grafts to achieve desired bone repair and regeneration.

Biocompatibility of Surface-Modified Biphasic Calcium Phosphate/Poly-L-Lactide Biocomposite in vitro and in vivo

The biocompatibility of surface-modified biphasic calcium phosphate （mBCP）/poly-L-Lactide （PLLA） biocomposite was investigated through a series of experiments in vitro and in vivo. Acute toxicity and short term systemic toxicity experiments revealed no toxicity of the materials. Hemolysis assay indicated the good blood compatibility of the composite. In cytotoxicity assay, L929 mouse fibroblasts could well differentiate and proliferate. Animal experiments in vivo were.performed by implanting the materials into rabbits muscle for 8 weeks. The decreasing of inflammatory cells, the building of fibrous tissue layer as well as the growing of blood cells into materials indicated the nontoxicity of the composite. Based on the experiment results, surfacemodified BCP/PLLA biocomposite is proven to have superior biocompatibility, which would be a promising bone repairing material.

One-step co-doping of ZnO and Zn^(2+)in osteoinductive calcium phosphate ceramics with synergistic antibacterial activity for regenerative repair of infected bone defect

How to endow bone grafts with long-term antibacterial activity and good bone regenerative ability to achieve the regenerative repair of infected bone defects has been the focus of the clinical treatment of osteomyelitis.The present study introduced a novel one-step route to realizing the co-doping of zinc oxide(ZnO)and zinc ion(Zn^(2+))in biphasic calcium phosphate(BCP)ceramics to utilize their synergistic antibacterial.Compared with the conventional BCP ceramics(BCP-Ca),the ZnO/Zn^(2+)co-doping ones(BCP-Zn)possessed strong antibacterial ability on E.coli and S.aureus as well as stimulated the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)effectively.The synergistic antibacterial mechanism of ZnO and Zn^(2+)was also investigated.BCP-Zn showed excellent osteoinductivity and angiogenesis at three months postoperatively in the canine intramuscular implantation model.Moreover,BCP-Zn exhibited excellent anti-infective ability and bone regenerative repair compared to BCP-Ca and control groups in the infected bone defect model of rat femur.Collectively,these findings suggest that the simultaneous introduction of ZnO/Zn^(2+)could have immense potential to expand the application of osteoinductive BCP ceramics in the regenerative repair of infected bone defects.

Preparation and properties of T-ZnO_(w) enhanced BCP scaffolds with double-layer structure by digital light processing

Bone scaffolds require both good bioactivity and mechanical properties to keep shape and promote bone repair.In this work,T-ZnO_(w) enhanced biphasic calcium phosphate(BCP)scaffolds with triply periodic minimal surface(TPMS)-based double-layer porous structure were fabricated by digital light processing(DLP)with high precision.Property of suspension was first discussed to obtain better printing quality.After sintering,T-ZnO_(w) reacts with b-tricalcium phosphate(β-TCP)to form Ca_(19)Zn_(2)(PO_(4))14,and inhibits the phase transition toα-TCP.With the content of T-ZnO_(w) increasing from 0 to 2 wt%,the flexural strength increases from 40.9 to 68.5 MPa because the four-needle whiskers can disperse stress,and have the effect of pulling out as well as fracture toughening.However,excessive whiskers will reduce the cure depth,and cause more printing defects,thus reducing the mechanical strength.Besides,T-ZnO_(w) accelerates the deposition of apatite,and the sample with 2 wt%T-ZnO_(w) shows the fastest mineralization rate.The good biocompatibility has been proved by cell proliferation test.Results confirmed that doping T-ZnO_(w) can improve the mechanical strength of BCP scaffolds,and keep good biological property,which provides a new strategy for better bone repair.

Osteoinduction by Ca-P biomaterials implanted into the muscles of mice

The osteoinduction of porous biphasic calcium phosphate ceramics (BCP) has been widely reported and documented,but little research has been performed on rodent animals,e.g.,mice.In this study,we report osteoinduction in a mouse model.Thirty mice were divided into two groups.BCP materials (Sample A) and control ceramics (Sample B) were implanted into the leg muscle,respectively.Five mice in each group were killed at 15,30,and 45 d after surgery.Sample A and Sample B were harvested and used for hematoxylin and eosin (HE) staining,immunohistochemistry (IHC) staining,and Alizarin Red S staining to check bone formation in the biomaterials.Histological analysis showed that no bone tissue was formed 15 d after implantation (0/5) in either of the two groups.Newly-formed bone tissues were observed in Sample A at 30 d (5/5) and 45 d (5/5) after implantation;the average amounts of newly-formed bone tissues were approximately 5.2% and 8.6%,respectively.However,we did not see any bone tissue in Sample B until 45 d after implantation.Bone-related molecular makers such as bone morphogenesis protein-2 (BMP-2),collagen type I,and osteopontin were detected by IHC staining in Sample A 30 d after implantation.In addition,the newly-formed bone was also confirmed by Alizarin Red S staining.Because this is the report of osteoinduction in the rodent animal on which all the biotechnologies were available,our results may contribute to further mechanism research.