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.

Experimental clearance rate and intraguild predation of jellyfish Cyanea nozakii

Cyanea nozakii,a common jellyfish distributed in offshore China,has a complex trophic relationship with other zooplankton groups.However,few studies have reported the predation rates and prey selection patterns of C.nozakii medusae on different prey items.Research is also lacking on the intraguild predation of Aurelia coerulea(another common bloom jellyfish in offshore China)by C.nozakii.To address the knowledge gaps,the clearance rates of C.nozakii for different prey items,including copepods(small<1000μm and large>1000μm),fish larvae,and gelatinous prey(hydromedusae,A.coerulea ephyrae,and chaetognaths),were measured.The influence of predator size on the clearance rate was also determined.Additionally,we examined the intraguild predation of C.nozakii on A.coerulea medusae.The clearance rates of C.nozakii varied widely with prey organisms,being independent of prey concentrations.Gelatinous organisms,except for chaetognaths,were captured with considerably high efficiency,followed by fish larvae and copepods,indicating the preferential prey selection of gelatinous organisms by C.nozakii.The clearance rate increased linearly with the cross-sectional area of C.nozakii.Body size in medusae may,to some extents,underpin their capacity to capture more prey by increasing the encounter rate and capture success through ontogeny.C.nozakii preyed voraciously on A.coerulea in high feeding efficiency,but the clearance rate decreased with increasing A.coerulea(as prey)size.This phenomenon of intraguild predation suggests a speculative hypothesis of potential population regulation of A.coerulea by C.nozakii.The information regarding the feeding ecology of C.nozakii reported in this study is important for understanding plankton dynamics in marine ecosystems with extensive occurrences of this jellyfish.

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.

Rice Grains from Slightly Saline Field Exhibited Unchanged Starch Physicochemical Properties but Enhanced Nutritional Values

This study aims to investigate grain quality and nutritional values of rice(Pokkali,a salt-tolerant cultivar;RD73,a new cultivar improved from KDML105 introgressed with Saltol QTL from Pokkali,and KDML105,a moderately salt-susceptible cultivar)grown under non-saline(0.04–0.87 dS/m)and slightly saline(1.08–4.83 dS/m)field conditions.The results revealed that salinity caused significant reduction in grain size but significant increments in reducing sugar and total protein contents in the grains.Nevertheless,the amounts of starch in the grains of KDML105 and Pokkali rice genotypes were unaffected by the stress.The starch granule size distribution was also unaffected by salinity.Interestingly,only starch from Pokkali was significantly diminished in amylose content,from 19.18%to 16.99%.Accordingly,parameters relating to starch gelatinization,retrogradation,and pasting properties of KDML105 and RD73 were unaffected by salinity;only Pokkali showed a significant increase in percentage of retrogradation along with a significant reduction in gelatinization enthalpy.In the saline field,total phenolic content and antioxidant capacity in the grains of all rice cultivars tended to increase,particularly in Pokkali.On average,essential element contents in grains from the saline-treated plants showed a 33%,32%,32%,22%,20%,11%,and 10%increase in total P,N,K,Mg,Zn,Fe,and Ca content,respectively.Interestingly,total Fe content exhibited the greatest percentage of increments in KDML105(187%).Taken together,cultivation of rice in the slightly saline field did not alter its eating and cooking qualities,while enhanced some nutritional properties such as proteins,minerals,and secondary metabolites like phenolic compounds.

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.

Effect of Butternut Squash (Cucurbita moschata) Seed Powder on the Chemical and Rheological Properties of Stirred Cultured Camel Milk and Yoghurt

Research shows that producing fermented camel milk is hard because of the milk’s inability to form a firm coagulum, attributed to low levels of κ-casein and ꞵ-lactoglobulin and the large casein micelle size, leading to a weak network of casein formation. In an effort to address this issue, researchers turned to corn starch as a thickening agent, discovering that a concentration of 2.0% effectively improved the viscosity and significantly reduced syneresis in stirred camel milk yoghurt and cultured camel milk. This study explores alternatives to corn starch, focusing on butternut squash seeds as a promising substitute due to their hydrocolloid composition. By incorporating butternut squash (Cucurbita moschata) seed powder (BSSP) as a thickening agent, this study aimed at enhancing the chemical and rheological properties of stirred camel milk yoghurt and cultured camel milk. Fermented camel milk was prepared using 4 litres of camel milk, 2% starter cultures (thermophilic culture for yoghurt and mesophilic aromatic culture for stirred cultured camel milk) and BSSP 0.0% (negative control), 0.4%, 0.8%, 1.2%, 1.6%, 2.0% mixed with 0.4% gelatin. 2.0% corn starch mixed with 0.4% gelatin was used as a standard for comparison. Results showed that increasing the BSSP level significantly (p < 0.05) decreased the moisture content while increasing the total solid content of stirred fermented camel milk products. There was an increase in ash content with an increase in BSSP levels. There was a significant (p < 0.05) reduction in the pH, with an increase in BSSP levels in stirred fermented camel milk samples. Increasing the concentration of BSSP from 0.4% to 2.0% resulted in a significant (p < 0.05) increase in viscosity and a reduction in syneresis of stirred camel milk yoghurt and stirred cultured camel milk samples. This study demonstrated that BSSP effectively enhances the viscosity, reduces syneresis and increases acidity in stirred fermented camel milk products during storage.

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.

电化学沉积法制备磷酸钙/明胶复合涂层的研究

采用脉冲电化学沉积法制备磷酸钙/明胶(CaP/Gelatin)复合涂层,对其进行表征及性能测试,旨在得到综合性能好的生物医用复合涂层.研究中讨论了不同脉冲电位、明胶浓度、沉积温度、电解液pH值等对复合涂层性能的影响.研究发现脉冲电位下涂层质量优于恒电位,XRD和FTIR分析表明复合涂层的成分为羟基磷灰石和明胶.SEM观察发现复合涂层是结构均匀的多孔网状或花瓣状微纳结构.复合涂层形貌及成分随脉冲电位、明胶浓度等实验条件的变换而变化.低电位利于复合涂层的结晶和沉积.改变明胶在钙磷电解液中的浓度,发现明胶浓度在0.1～1.0 g/L范围内不影响涂层形貌,但影响明胶在复合涂层中的含量.沉积温度为50℃,电解液pH值在明胶等电点以下保证了复合涂层具有较好的结晶性、均匀性.AlamarBlue检测表明复合涂层具有较好的生物相容性.

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.

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.

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.

THE STRUCTURE OF THERMOPLASTIC STARCH

The granular structure, crystal structure and gelatinization temp. of thermoplastic starch were studied with a polarized light microscope and a scanning electron microscope, and the crystallinity and crystalline patterns were determined through X ray diffraction. The results indicate that the original granular structure and spherical crystalline structure of starch were disrupted by the action of pressure, heat and shear force with the help of additives. The starch can be melted during extrusion, and part of the spheric crystal was destroyed and changed into a continual amorphous with a few crystalline fractions dispersed in it. The configuration of starch molecules changed from double helices to single helix, which indicated the formation of the complex.

静电纺羟基磷灰石/明胶微纳复合纤维

本文从仿生角度出发,模拟细胞外基质独特结构,采用静电纺丝法成功制备出HA均匀分布的HA/Gelatin复合纤维。根据影响静电纺丝的主要因素,分别考察了聚合物浓度、无机物含量、溶剂浓度、电纺电压等因素对纤维形貌和结构的影响。研究结果表明:聚合物浓度是制备复合纤维的首要影响因素,影响复合纤维的直径;无机物的添加使聚合物中的氢键减少,降低了电纺液的粘度,影响复合纤维中珠状物的形成;制备分布均匀的电纺纤维,溶剂起很大的作用,影响纤维的粘联;电纺电压增大使电场力过大,聚合物被强力拉伸,单根纤维出现卷曲。

Gelatin／BaTiO3核壳复合粒子的制备及电场响应性能研究

以平均粒径为0．7～0．9μm的明胶（Gelatin）微球为基核，通过定向沉积自组装法成功制各了以Gelatin为核、BaTiO3为壳的Gelatin／BaTiO3核壳复合粒子。利用TEM、XRD、FT-IR、TG和光学接触角测量等技术对复合粒子的形貌、结构、组成及表面亲水性能进行了研究。结果表明，复合粒子为球形微粒，具有良好的表面亲水性和分散稳定性，BaTiO3壳层约占粒子总质量的18．8％，具有立方相晶相结构。将粒子分散到水凝胶弹性体中，测量在有／无电场作用下得到的弹性体的储能模量，借以考察复合粒子在弹性体中的电场响应性能，发现Gelatin／BaTiO3复合粒子的电场响应性能明显强于纯BaTiO3粒子。说明将BaTiO3包覆在聚合物上能显著提高BaTiO3粒子的电场响应性能。

质粒修饰BMSC联合纳米支架促进软骨缺损修复的相关研究

目的探究慢病毒介导聚蛋白多糖(Aggrecan)过表达质粒转染骨髓间充质干细胞联合Gelatin/PLGA纳米纤维多孔支架向软骨细胞转化及软骨缺损修复的作用效果。方法构建慢病毒聚蛋白多糖过表达载体,培养骨髓间充质干细胞,构建兔模型软骨缺损模型,实验组造模后植入Gelatin/PLGA三维支架+转染聚蛋白多糖过表达载体骨髓间充质干细胞复合物;阴性对照组造模后植入Gelatin/PLGA三维支架+未转染骨髓间充质干细胞复合物;模型组造模后加入生理盐水处理。体外实验利用HE染色和免疫组织化学染色检测骨髓间充质干细胞联合Gelatin/PLGA纳米纤维多孔支架的联合培养体系向软骨细胞转化中聚蛋白多糖和Ⅱ型胶原蛋白的表达。构建兔膝关节软骨缺损模型,HE染色和免疫组织化学分析复合支架材料对软骨缺损的修复效果。结果 a)实验组(Gelatin/PLGA纳米纤维多孔支架+转染Aggrecan过表达载体骨髓间充质干细胞复合物)中支架表面黏附的细胞与对照组(Gelatin/PLGA纳米纤维多孔支架+未转染骨髓间充质干细胞)相比,细胞数明显增多;b)动物模型大体观察结果显示,实验组的修复效果要明显优于阴性对照组;c)动物模型HE染色结果显示,阴性对照组多为纤维性修复,而实验组多为细胞性修复;d)免疫组织化学染色结果显示,实验组修复软骨组织中Aggrecan和Ⅱ型胶原含量要明显优于阴性对照组。结论 Gelatin/PLGA三维支架加转染Aggrecan过表达载体骨髓间充质干细胞复合物可以促进BMSC向软骨细胞的分化,并且分泌更多的含Aggrecan和Ⅱ型胶原成分的细胞外基质,从而发挥其促进软骨缺损修复的作用。