One-step cell biomanufacturing platform:porous gelatin microcarrier beads promote human embryonic stem cell-derived midbrain dopaminergic progenitor cell differentiation in vitro and survival after transplantation in vivo

Numerous studies have shown that cell replacement therapy can replenish lost cells and rebuild neural circuitry in animal models of Parkinson’s disease.Transplantation of midbrain dopaminergic progenitor cells is a promising treatment for Parkinson’s disease.However,transplanted cells can be injured by mechanical damage during handling and by changes in the transplantation niche.Here,we developed a one-step biomanufacturing platform that uses small-aperture gelatin microcarriers to produce beads carrying midbrain dopaminergic progenitor cells.These beads allow midbrain dopaminergic progenitor cell differentiation and cryopreservation without digestion,effectively maintaining axonal integrity in vitro.Importantly,midbrain dopaminergic progenitor cell bead grafts showed increased survival and only mild immunoreactivity in vivo compared with suspended midbrain dopaminergic progenitor cell grafts.Overall,our findings show that these midbrain dopaminergic progenitor cell beads enhance the effectiveness of neuronal cell transplantation.

Three-dimensional porous bimetallic metal–organic framework/gelatin aerogels: A readily recyclable peroxymonosulfate activator for efficient and continuous organic dye removal

As promising catalysts for the degradation of organic pollutants,metal–organic frameworks(MOFs)often face limitations due to the particle agglomeration and challenging recovery in liquid-catalysis application,stemming from their powdery nature.Engineering macroscopic structures from pulverous MOF is thus of great importance for broadening their practical applications.In this study,three-dimensional porous MOF aerogel catalysts were successfully fabricated for degrading organic dyes by activating peroxymonosulfate(PMS).MOF/gelatin aerogel(MOF/GA)catalysts were prepared by directly integrating bimetallic FeCo-BDC with gelatin solutions,followed by freeze-drying and low-temperature calcination.The FeCo-BDC-0.15/GA/PMS system exhibited remarkable performance in degrading various organic dyes,eliminating 99.2%of rhodamine B within a mere 5 min.Compared to the GA/PMS system,there was over a 300-fold increase in the reaction rate constant.Remarkably,high removal efficiency was maintained across varying conditions,including different solution pH,co-existing inorganic anions,and natural water matrices.Radical trapping experiments and electron paramagnetic resonance analysis revealed that the degradation involved radical(SO_(4)^(-)·)and non-radical routes(^(1)O_(2)),of which ^(1)O_(2) was dominant.Furthermore,even after a continuous 400-min reaction in a fixed-bed reactor at a liquid hourly space velocity of 27 h^(-1),the FeCo-BDC/GA composite sustained a degradation efficiency exceeding 98.7%.This work presents highly active MOF-gelatin aerogels for dye degradation and expands the potential for their large-scale,continuous treatment application in organic dye wastewater management.

Bio-inspired Hydroxyapatite/Gelatin Transparent Nanocomposites

Hydroxyapatite(HA)nanoparticles impart outstanding mechanical properties to organicinorganic nanocomposites in bone.Inspired by the composite structure of HA nanoparticles and collagen in bone,a high performance HA/gelatin nanocomposite was first developed.The nanocomposites have much better mechanical properties(elongation at break 29.9%,tensile strength 90.7 MPa,Young’s modulus 5.24 GPa)than pure gelatin films(elongation at break 9.3%,tensile strength 90.8 MPa,Young’s modulus 2.5 GPa).In addition,the composite films keep a high transmittance in visible wavelength range from 0%to 60%of the HA solid content.These differences in properties are attributed to the homogeneous distribution of HA nanoparticles in the gelatin polymer matrix and the strong interaction between the particle surfaces and the gelatin molecules.This protocol should be promising for HA-based nanocomposites with enhanced mechanical properties for biomedical applications.

Constructing a biofunctionalized 3D-printed gelatin/sodium alginate/chitosan tri-polymer complex scaffold with improvised biological andmechanical properties for bone-tissue engineering

Sodium alginate(SA)/chitosan(CH)polyelectrolyte scaffold is a suitable substrate for tissue-engineering application.The present study deals with further improvement in the tensile strength and biological properties of this type of scaffold to make it a potential template for bone-tissue regeneration.We experimented with adding 0%–15%(volume fraction)gelatin(GE),a protein-based biopolymer known to promote cell adhesion,proliferation,and differentiation.The resulting tri-polymer complex was used as bioink to fabricate SA/CH/GEmatrices by three-dimensional(3D)printing.Morphological studies using scanning electron microscopy revealed the microfibrous porous architecture of all the structures,which had a pore size range of 383–419μm.X-ray diffraction and Fourier-transform infrared spectroscopy analyses revealed the amorphous nature of the scaffold and the strong electrostatic interactions among the functional groups of the polymers,thereby forming polyelectrolyte complexes which were found to improve mechanical properties and structural stability.The scaffolds exhibited a desirable degradation rate,controlled swelling,and hydrophilic characteristics which are favorable for bone-tissue engineering.The tensile strength improved from(386±15)to(693±15)kPa due to the increased stiffness of SA/CH scaffolds upon addition of gelatin.The enhanced protein adsorption and in vitro bioactivity(forming an apatite layer)confirmed the ability of the SA/CH/GE scaffold to offer higher cellular adhesion and a bone-like environment to cells during the process of tissue regeneration.In vitro biological evaluation including the MTT assay,confocal microscopy analysis,and alizarin red S assay showed a significant increase in cell attachment,cell viability,and cell proliferation,which further improved biomineralization over the scaffold surface.In addition,SA/CH containing 15%gelatin designated as SA/CH/GE15 showed superior performance to the other fabricated 3D structures,demonstrating its potential for use in bone-tissue engineering.

Fish Gelatin-Based Film Containing Maillard Reaction Products:Properties and Its Use as Bag for Packing Chicken Skin Oil

Maillard reaction is a non-enzymatic browning reaction and its products(MRPs)have been proven to possess antioxidant properties.This research aimed to produce a fish gelatin-based packaging incorporated with MRPs to retard lipid oxidation in chicken skin oil(CSO)during storage at ambient temperature(28℃–30℃).MRPs produced from fish gelatin and fructose(1:1,90℃,pH 11)showed the highest antioxidant properties compared to those prepared under other conditions.Different glycerol/MRPs ratios(30:0,25:5,20:10,15:15,10:20,5:25,0:30)were incorporated into the film and resulting films were characterized.Glycerol/MRPs at 10:20 ratio was chosen to add into the film prior to bag preparation via heat sealing method.CSO packed in the bag was monitored for lipid hydrolysis and oxidation during 15 days of storage(30℃±0.5℃,RH 52%±5%).After 15 days,quality deterioration was lower in CSO packed in the prepared gelatin bag as evidenced by lower FFA,TBARS,and volatile compounds in comparison with CSO packed in LDPE bag.Fish gelatin film added with MRPs possessed an excellent water vapor barrier property(WV-BP).This finding indicated that MRPs could be used to substitute glycerol and simultaneously could serve as antioxidants for the developed active bag.The novel packaging can be a potential alternative packaging for retarding lipid oxidation of lipid or fatty foods.

Studies of XRD and FTIR on Synthesized Novel Hybrid Thin Film Made of Hydroxyapatite, Poly Vinyl Alcohol and Gelatin for Biomedical Application

With the modern advancement of treatment approaches in medical science, the application of biomaterials in tissue engineering provides a remarkable opportunity to overcome graft rejection as well as proper wound healing. In this study, novel hybrid films have been synthesized by incorporation of polyvinyl alcohol (PVA), gelatin, and gelatin with glycerin along with different concentrations of pre-prepared hydroxyapatite (HAP) by solution casting method at room temperature in a biosafety cabinet. Glutaraldehyde has been added as a crosslinker in this whole procedure. Fourier-transform infrared spectroscopy (FTIR), X-Ray Diffraction (XRD) have been conducted to observe and compare the structural and chemical stability of the synthesized hybrid film properties. The FTIR results and X-Ray Diffraction analyses confirmed the chemical interactions between HAP, PVA, gelatin, and glycerin have occurred. The crystallinity of HAP also remains in all the prepared hybrid film samples that are observed in XRD. It is expected that these newly synthesized hybrid films could be a better opportunity for various sectors of tissue engineering such as skin, bone, tendon, and cartilage. These synthesized hybrid films can be suitable for wound healing covering. These studies could be a new scope for long-term drug delivery directly on wound sites in diabetic gangrene foot or burn patients as well as cartilage or joint replacement therapy.

Development and Characterization of Hybrid Film Made of Hydroxyapatite, Poly Vinyl Alcohol and Gelatin for Biomedical Application

In the recent research field of bone tissue engineering, polymeric materials play an implacable role in mimes the natural behavior of hard and soft tissues. In some medical conditions such as diabetics, osteoarthritis, burns, or joint replacement conditions, this polymeric materials implication enhances the internal mechanical activities which result in the early recovery of disease by facilitating the wound healing process. In this study, hybrid films have been synthesized based on polyvinyl alcohol (PVA), gelatin, and gelatin with glycerin incorporated with different concentrations of pre-prepared hydroxyapatite (HAP) by solution casting method at room temperature in biosafety cabinet. Glutaraldehyde has been added as a crosslinker in this whole procedure. The mechanical property, swelling, and porosity percentage have been conducted to characterize the structural stability of the synthesized hybrid films. Porosity and swelling of samples are also represented by proper biocompatibility (>90% porosity and swelling in DDW and PBF vary between 287%~72%). Tensile strength (TS), E modulus (Young’s modulus), Elongation at maximum, and Elongation at break are observed to perceive the mechanical properties of hybrid film samples, which are compatible with mechanical properties of different tissue such as trabecular bone, articular cartilage, tendon, nerve and skin tissue. Though, biocompatibility tests both in vivo and in vitro are essential for clinical application in the future. However, the experiment carried out till now explains the true possibility of newly synthesized hybrid films for long-term drug delivery directly on wound sites for wound healing and burn dressing patients in head-neck surgery reconstruction, diabetic gangrene foot, as well as cartilage or joint replacement therapy.

Tissue Regeneration in Infected Wounds of Albino Rats Using Ciprofloxacin-Loaded Gelatin Microspheres Incorporated Collagen Scaffold: A Histological Approach with H&E Staining

A wound care system consisting of ciprofloxacin-loaded gelatin microspheres impregnated in a macroporous collagen scaffold was created to effectively control wound infection and regenerate soft tissue at the wound site.Histological and biochemical alterations were observed in infected wounds treated with these scaffolds in Albino Wistar rats.Furthermore,the study examined the immediate and prolonged release of ciprofloxacin from the scaffolds,as well as their function in eliminating bacterial infections and expediting the process of skin healing and regeneration.The developed technique was followed in the streamlined process of creating these collagen scaffolds.Compared to untreated wounds,the group receiving scaffold treatment experienced a faster rate of wound closure.It was noted that the rate of infections was considerably reduced and that full soft tissue regeneration occurred within 12 days.The development of well-deposited collagen bundles in the treated groups was demonstrated by H&E staining,which verified the flawless regeneration of the dermis and epidermis.The antimicrobial agent-loaded gelatin microspheres impregnated into the porous collagen scaffold demonstrated remarkable soft tissue regeneration and efficient infection control at the wound site.

Antibacterial Chitosan-Gelatin Microcapsules Modified with Green-Synthesized Silver Nanoparticles for Food Packaging

Silver nanoparticles(Ag NPs)are an effective antibacterial agent,but their application in food packaging is limited due to their easy agglomeration and oxidation.In this study,antibacterial microcapsules were fabricated using Ginkgo biloba essential oil(GBEO)as core material and chitosan and type B gelatin biopolymer as capsule mate-rials.These antibacterial microcapsules were then modified with green-synthesized Ag NPs,blended into the bio-polymer polylactic acid(PLA),and finally formed as films.Physicochemical properties and antibacterial activity against Escherichia coli(E.coli)and Staphylococcus aureus(S.aureus)were evaluated.Results showed that the prepared antibacterial PLA films exhibited excellent antibacterial activity against foodborne pathogens.Its TVC exceeded the limit value of 7 log CFU/g at 7 days compared with the 5 days of pure PLA films.Therefore,these films can extend the shelf life of grass carp fillets by 2–3 days under refrigeration.

3D-printed engineered bacteria-laden gelatin/sodium alginate composite hydrogels for biological detection of ionizing radiation

Nuclear safety is a global growing concern,where ionizing radiation(IR)is a major injury factor resulting in serious damage to organisms.The detection of IR is usually conducted with physical dosimeters;however,biological IR detection methods are deficient.Here,a living composite hydrogel consisting of engineered bacteria and gelatin/sodium alginate was 3D-printed for the biological detection of IR.Three strains of PrecA::egfp gene circuit-containing engineered Escherichia coli were constructed with IR-dependent fluorescence,and the DH5αstrain was finally selected due to its highest radiation response and fluorescence.Engineered bacteria were loaded in a series of gelatin/sodium alginate matrix hydrogels with different rheology,3D printability and bacterial applicability.A high-gelatin-content hydrogel containing 10%gelatin/1.25%sodium alginatewas optimal.The optimal living composite hydrogelwas 3D-printedwith the special bioink,which reported significant green fluorescence underγ-ray radiation.The living composite hydrogel provides a biological strategy for the detection of environmental ionizing radiation.

Alginate/gelatin/boron-doped hydroxyapatite-coated Ti implants:in vitro and in vivo evaluation of osseointegration

In this study,boron-doped hydroxyapatite(BHT)-loaded alginate/gelatin-based(A/G)hydrogel coating on Ti was fabricated to support bone integration through triggering osteoinduction,vascularization and immunomodulation.Initially,highly reproducible,cheap and time-effective BHT was produced,which significantly promoted higher osteogenic and angiogenic maturation,while a mild innate immune response was observed.The immense potential of BHT was evidenced by the production of a gap-filling A/G/BHT interphase on Ti implants to mimic the osseous extracellular matrix to achieve functional bridging and exert control over the course of innate immune response.We initially aminosilanized the implant surface using 3-aminopropyl triethoxysilane,and then coated it with 0.25%w/v alginate with 20 mM 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide to allowthe A/G/BHT pre-gel to disperse evenly and covalently attach on the surface.The pre-gel was added with 0.2 M NaCl to homogeneously blend BHT in the structure without inducing ionic crosslinking.Then,the coated implants were freeze-dried and stored.The coated layer demonstrated high cohesive and adhesive strength,and 8-month-long shelf-life at room temperature and normal humidity.The A/G/BHT was able to coat an irregularly shaped Ti implant.Osteoblasts and endothelial cells thrived on the A/G/BHT,and it demonstrated greatly improved osteogenic and angiogenic capacity.Moreover,A/G/BHT maintained macrophage viability and generated an acute increase in immune response that could be resolved rapidly.Finally,A/G/BHT was shown to induce the robust integration of implant in a rabbit femur osteochondral model within 2months.Therefore,we concluded that A/G/BHT coatings could serve as amultifunctional reservoir,promoting the strong and rapid osseointegration of metallic implants.

Comparison of ethanol-soaked gelatin sponge and microspheres for hepatic arterioportal fistulas embolization in hepatic cellular carcinoma

Hepatic arterioportal fistulas(APFs)are common in hepatocellular carcinoma(HCC).Moreover,correlated with poor prognosis,APFs often complicate antitumor treatments,including transarterial chemoembolization(TACE).AIM To compare the efficacy of ethanol-soaked gelatin sponges(ESG)and microspheres in the management of APFs and their impact on the prognosis of HCC.METHODS Data from patients diagnosed with HCC or hepatic APFs between June 2016 and December 2019 were retrospectively analyzed.Furthermore,APFs were embolized with ESG(group E)or microspheres(group M)during TACE.The primary outcomes were disease control rate(DCR)and objective response rate(ORR).The secondary outcomes included immediate and first follow-up APF improvement,overall survival(OS),and progression-free survival(PFS).RESULTS Altogether,91 participants were enrolled in the study,comprising 46 in group E and 45 in group M.The DCR was 93.5%and 91.1%in groups E and M,respectively(P=0.714).The ORRs were 91.3%and 66.7%in groups E and M,respectively(P=0.004).The APFs improved immediately after the procedure in 43(93.5%)patients in group E and 40(88.9%)patients in group M(P=0.485).After 2 mo,APF improvement was achieved in 37(80.4%)and 33(73.3%)participants in groups E and M,respectively(P=0.421).The OS was 26.2±1.4 and 20.6±1.1 mo in groups E and M,respectively(P=0.004),whereas the PFS was 16.6±1.0 and 13.8±0.7 mo in groups E and M,respectively(P=0.012).CONCLUSION Compared with microspheres,ESG embolization demonstrated a higher ORR and longer OS and PFS in patients of HCC with hepatic APFs.

Enhanced wound healing and hemostasis with exosome-loaded gelatin sponges from human umbilical cord mesenchymal stem cells

BACKGROUND Rapid wound healing remains a pressing clinical challenge,necessitating studies to hasten this process.A promising approach involves the utilization of human umbilical cord mesenchymal stem cells(hUC-MSCs)derived exosomes.The hypothesis of this study was that these exosomes,when loaded onto a gelatin sponge,a common hemostatic material,would enhance hemostasis and accelerate wound healing.AIM To investigate the hemostatic and wound healing efficacy of gelatin sponges loaded with hUC-MSCs-derived exosomes.METHODS Ultracentrifugation was used to extract exosomes from hUC-MSCs.Nanoparticle tracking analysis(NTA),transmission electron microscopy(TEM),and western blot techniques were used to validate the exosomes.In vitro experiments were performed using L929 cells to evaluate the cytotoxicity of the exosomes and their impact on cell growth and survival.New Zealand rabbits were used for skin irritation experiments to assess whether they caused adverse skin reactions.Hemolysis test was conducted using a 2%rabbit red blood cell suspension to detect whether they caused hemolysis.Moreover,in vivo experiments were carried out by implanting a gelatin sponge loaded with exosomes subcutaneously in Sprague-Dawley(SD)rats to perform biocompatibility tests.In addition,coagulation index test was conducted to evaluate their impact on blood coagulation.Meanwhile,SD rat liver defect hemostasis model and full-thickness skin defect model were used to study whether the gelatin sponge loaded with exosomes effectively stopped bleeding and promoted wound healing.RESULTS The NTA,TEM,and western blot experimental results confirmed that exosomes were successfully isolated from hUC-MSCs.The gelatin sponge loaded with exosomes did not exhibit significant cell toxicity,skin irritation,or hemolysis,and they demonstrated good compatibility in SD rats.Additionally,the effectiveness of the gelatin sponge loaded with exosomes in hemostasis and wound healing was validated.The results of the coagulation index experiment indicated that the gelatin sponge loaded with exosomes had significantly better coagulation effect compared to the regular gelatin sponge,and they showed excellent hemostatic performance in a liver defect hemostasis model.Finally,the full-thickness skin defect healing experiment results showed significant improvement in the healing process of wounds treated with the gelatin sponge loaded with exosomes compared to other groups.CONCLUSION Collectively,the gelatin sponge loaded with hUC-MSCs-derived exosomes is safe and efficacious for promoting hemostasis and accelerating wound healing,warranting further clinical application.

Development of Gelatin-Based Active Packaging and Its Application in Bread Preservation

The issue of plastic pollution has attracted widespread social attention.Gelatin is valued as a degradable bio-based material,especially as an edible active packaging material.However,the commonly used solution-casting filmforming technology limits the mass production of gelatin films.In order to improve the production efficiency and enhance the commercial value of gelatin films,in this study,fish gelatin(FG)particles were successfully blended with essential oils(EOs)to prepare active films by melt extrusion technique,a common method for commercial plastics,and applied to bread preservation.FG and EOs showed good compatibility with each other.The elongation at break was enhanced in all samples of films containing EOs.The addition of EOs weakened the oxygen barrier properties of the FG films.The gelatin films containing clove EO showed the highest DPPH radical scavenging rate of 39.38%.The films containing oregano EO showed the highest antibacterial activity,with 98.84%and 99.86% against S.aureus and E.coli,respectively.The bread preservation results showed a slower microbial growth rate in the samples preserved by the active films.The bread samples preserved in commercial PE film showed mold on day 3,and the bread samples preserved in active gelatin film showed mold visible to the naked eye by day 7.This study demonstrates the potential of gelatin-based activated films for commercial application in baked goods preservation.

Broiler Feet Gelatine

The two waves period of the COVID-19 pandemic saw the use of hydro- alcoholic gel and the consumption of capsules containing improved traditional remedies. At one point, there was a stock-out and a price increase forthese products. Furthermore, in the food industry, the catering industry adopts gelatin in its current practice. Pig gelatin dominates the international market. And for some religious practices, pork is forbidden and yet these people consume them without taking notice. The production of gelatin from broiler feet seems economically viable because broiler feet are considered slaughterhouse waste that is sold at very low prices. The poultry industry has seen an increase in broiler farming over the last twenty years. However, the latter has all the characteristics required for the production of gelatin. It will therefore comply with the standards of use described in the international codex oenological for gelatins. Physical and chemical analyses such as, ash content, moisture content, and pH measurements were done for the extracted gelatins. Sensible elements are checked with ED XRF spectroscopy. All the results were good and showed without any doubt that broiler gelatin is edible.

Studies on Adriamycin Magnetic Gelatin Microspheres

The preparation and properties of adriamycin magnetic gelatin microspheres(Adr- MG-ms)were reported.The synthesis of magnetic iron oxide ultrafine particle and embolization effects of magnetic gelatin microspheres(MG-ms)in dog were studied.Adr- MG-ms consist of 2%(w/w)of adriamycin(Adr)as the core,and 68% of gelatin and 30% of magnetite as the shell with a mean particle size of 22 μm. In vitro experiment,the release rate of drug demonstrated that the microspheres have sustained-release properties.The average diameter of magnetic iron oxide was approximately l0 nm. Transcatheter embolization with MG-ms and ￣(99m)Tc-labelled MG-ms was performed under external magenet control in dog liver,respectively.Gamma photography and angiogram revealed that MG-ms level was almost equal both in left and right hepatic arteries without magnet,while with magnet(1200 Gs),MG-ms level in left hepatic artery(target site)was about 2.25 fold higher than in right hepatic artery,and few MG-ms in thyroid gland,brain and heart was observed.Results showed that the MG-ms is a promising embolic agent for treatment of hepatic cancer under external magnet control.

BaTiO3/gelatin核壳复合粒子的制备及电场响应性能研究

为了提高BaTiO3颗粒在含水弹性体中的电场响应能力,以粒径约为500nm的单分散球型Ba-TiO3颗粒为基础,采用微乳液法成功制备了BaTiO3为核、明胶(gelatin)为壳的BaTiO3/gelatin核壳复合粒子。利用TEM、XRD、FT-IR、TG和光学接触角测量等手段对粒子的组成、结构及表面亲水性能进行了表征和测量。结果表明,在立方相BaTiO3核的表面均匀包覆了1层约40～50nm的gelatin,Gelatin壳层约占粒子总重的4.0%,且粒子表面保持良好的亲水性。将粒子分散到含水胶体体系中,通过测量在无/有电场作用下胶凝得到的含水弹性体的储能模量考察了粒子在此体系中的电场响应性能,发现BaTiO3/gelat-in复合粒子在明胶(或壳聚糖)水凝胶弹性体中的电场响应性能均明显强于纯BaTiO3粒子,而且在明胶水凝胶中的响应性能更强。结果说明,聚合物包覆能够明显提高BaTiO3粒子的电场响应性能,而且粒子表面与分散体系相容性越高,粒子的响应性能越好。

Extraction Characteristics of Anthocyanins from Preliminarily-processed Products of Purple Sweet Potatoes during the Gelatinization Process

Objective] This study aimed to investigate the method for efficient utilization and development of purple sweet potatoes. [Method] Purple sweet potatoes were dried at two specific temperatures and prepared into preliminarily-processed products for gelatinization simulation to analyze the extraction amount of anthocyanins from gelatinized samples at different gelatinization stages. [Result] During the gelatinization process, the extraction rate of anthocyanins from purple sweet potato samples reached the highest as the temperature rised from 90 ℃ to 95 ℃,and the extraction amount of anthocyanins reached the maximum at 15 min postheat preservation at 95 ℃. Purple sweet potato samples dried at 60 ℃ exhibited larger retention amount, larger maximum extraction amount and higher maximum extraction rate of anthocyanins compared with those dried at 110 ℃. [Conclusion] Drying at low temperatures and appropriately shortening the initial gelatinization stage below 90 ℃ is conducive to the retention and extraction of anthocyanins from purple sweet potatoes.

STUDY ON HYDROLYSIS OF MACROMOLECULAR GELATIN WITH ENZYMES IN COMBINATION MODE

The enzymatic hydrolysis of macromolecular gelatin with AS1.398 neutral protease, bromelain and their combinations was studied by estimating the molecular weights of their hydrolytic products. It was discovered that the products hydrolyzed by using combination enzymes had lower molecular weight than those obtained by using single ones, and the kind of enzymes, their combination mode and addition sequence are effective ways to control the molecular weights of gelatin hydrolyzates.

Effect of porous titanium coated with IGF-1 and TGF-β_1 loaded gelatin microsphere on function of MG63 cells

Porous titanium with porosity of 60% was prepared by metal injection molding（MIM）,and coated with gelatin sustained-release microspheres which were made by improved emulsified cold condensation method.The effects of porous titanium coated with insulin-like growth factor-1（IGF-1） and transforming growth factor-β1（TGF-β1） gelatin microspheres on the function of MG63 cells were evaluated in vitro.The results show that porous titanium coated with gelatin sustained-release microspheres has no cytotoxicity.The IGF-1 and TGF-β1 loading concentrations are positively correlative with the proliferation and differentiation of MG63 after co-culturing with the concentrations of IGF-1 and TGF-β1 gelatin microspheres in the range of 0.1-10 ng/mg and 0.25-2.5 ng/mg,respectively.The MG63 cells exhibit the best proliferation and differentiation with the IGF-1 and TGF-β1 loading concentrations of 10 ng/mg and 2.5 ng/mg,respectively.The joint application of IGF-1 and TGF-β1 group,which promote adhesion,proliferation and differentiation of MG63 cells,is superior to a single application group.