Biocompatibility properties of composite scaffolds based on 1,4-butanediamine modified poly(lactide-co-glycolide) and nanobioceramics

Three-dimensional biodegradable porous scaffolds play an important role in tissue engineering. The degradable scaffold material, based on 1,4-butanediamine-modified poly(lactide-co-glycolide) (BMPLGA), nano-bioactive glass (NBAG), and nano-β-tricalciumphosphate (β-TCP), was prepared by a solution-casting/salt-leaching method. The biological properties were studied by using cell cytotoxicity, von Kossa staining, alkaline phosphatase activity, hemolytic test, acute toxicity, and genetic toxicity test. The MTT results indicated that the BMPLGA/NBAG-β-TCP materials did not show any cytotoxicity. The result of von Kossa staining showed that the introduction of the NBAG and β-TCP promoted fibroblastic differentiation and improved the mineral deposition of the BMPLGA matrix. In addition, the presence of NBAG and β-TCP in the composite further enhanced the ALP activity in comparison with the sole BMPLGA material. The hemolytic potential showed that the nanocomposite scaffolds were non-hemolytic. The BMPLGA/NBAG-β-TCP scaffolds showed no acute systemic toxicity or mutagenic action. Therefore, the results indicated the BMPLGA/NBAG-β-TCP nanocomposite scaffold could be considered as a potential bone tissue engineering implant.     

Incorporation of nanohydroxyapatite and vitamin D3 into electrospun PCL/Gelatin scaffolds: The influence on the physical and chemical properties and cell behavior for bone tissue engineering

Scaffold, an essential element of tissue engineering, should provide proper physical and chemical properties and evolve suitable cell behavior for tissue regeneration. Polycaprolactone/Gelatin (PCL/Gel)‐based nanocomposite scaffolds containing hydroxyapatite nanoparticles (nHA) and vitamin D3 (Vit D3) were fabricated using the electrospinning method. Structural and mechanical properties of the scaffold were determined by scanning electron microscopy (SEM) and tensile measurement. In this study, smooth and bead‐free morphology with a uniform fiber diameter and optimal porosity level with appropriate pore size was observed for PCL/Gel/nHA nanocomposite scaffold. The results indicated that adding nHA to PCL/Gel caused an increase of the mechanical properties of scaffold. In addition, chemical interactions between PCL, gelatin, and nHA molecules were shown with XRD and FT‐IR in the composite scaffolds. MG‐63 cell line has been cultured on the fabricated composite scaffolds; the results of viability and adhesion of cells on the scaffolds have been confirmed using MTT and SEM analysis methods. Here in this study, the culture of the osteoblast cells on the scaffolds showed that the addition of Vit D3 to PCL/Gel/nHA scaffold caused further attachment and proliferation of the cells. Moreover, DAPI staining results showed that the presence and viability of the cells were greater in PCL/Gel/nHA/Vit D3 scaffold than in PCL/Gel/nHA and PCL/Gel scaffolds. The results also approved increasing cell proliferation and alkaline phosphatase (ALP) activity for MG‐63 cells cultured on PCL/Gel/nHA/Vit D3 scaffold. The results indicated superior properties of hydroxyapatite nanoparticles and vitamin D3 incorporated in PCL/Gel scaffold for use in bone tissue engineering.     

Oxygen Plasma Technology-Assisted Preparation of Three-Dimensional Reduced Graphene Oxide/Polypyrrole/Strontium Composite Scaffold for Repair of Bone Defects Caused by Osteoporosis

Repairs of bone defects caused by osteoporosis have always relied on bone tissue engineering. However, the preparation of composite tissue engineering scaffolds with a three-dimensional (3D) macroporous structure poses huge challenges in achieving osteoconduction and osteoinduction for repairing bone defects caused by osteoporosis. In the current study, a three-dimensional macroporous (150–300 μm) reduced graphene oxide/polypyrrole composite scaffold modified by strontium (Sr) (3D rGO/PPY/Sr) was successfully prepared using the oxygen plasma technology-assisted method, which is simple, safe, and inexpensive. The findings of the MTT assay and AO/EB fluorescence double staining showed that 3D rGO/PPY/Sr has a good biocompatibility and effectively promoted MC3T3-E1 cell proliferation. Furthermore, the ALP assay and alizarin red staining showed that 3D rGO/PPY/Sr increased the expression levels of ALP activity and the formation of calcified nodules. The desirable biocompatibility, osteoconduction, and osteoinduction abilities, assure that the 3D macroporous rGO/PPY/Sr composite scaffold offers promising potential for use in the repair of bone defects caused by osteoporosis in bone tissue engineering.     

Preparation,Characterization, and In Vitro Biological Evaluation of PLGA/Nano-Fluorohydroxyapatite (FHA) Microsphere-Sintered Scaffolds for Biomedical Applications

In this research, the novel three-dimensional (3D) porous scaffolds made of poly(lactic-co-glycolic acid) (PLGA)/nano-fluorohydroxyapatite (FHA) composite microspheres was prepared and characterize for potential bone repair applications. We employed a microsphere sintering method to produce 3D PLGA/nano-FHA scaffolds composite microspheres. The mechanical properties, pore size, and porosity of the composite scaffolds were controlled by varying parameters, such as sintering temperature, sintering time, and PLGA/nano-FHA ratio. The experimental results showed that the PLGA/nano-FHA (4:1) scaffold sintered at 90 °C for 2 h demonstrated the highest mechanical properties and an appropriate pore structure for bone tissue engineering applications. Furthermore, MTT assay and alkaline phosphatase activity (ALP activity) results ascertained that a general trend of increasing in cell viability was seen for PLGA/nano-FHA (4:1) scaffold sintered at 90 °C for 2 h by time with compared to control group. Eventually, obtained experimental results demonstrated PLGA/nano-FHA microsphere-sintered scaffold deserve attention utilizing for bone tissue engineering.     

纳米羟基磷灰石/丝素蛋白复合支架材料的降解特性及生物相容性研究

应用共混法制备了纳米羟基磷灰石/丝素蛋白复合支架材料, 通过体外降解和细胞培养实验研究了复合支架材料的降解特性和生物相容性. 体外降解实验结果显示, 复合支架材料具有稳定的降解能力; 在降解过程中, 羟基磷灰石由于与降解液发生钙、磷等离子的交换, 使其结晶得到了进一步生长和完善. 利用细胞计数法、四甲基偶氮唑盐(MTT)比色法和碱性磷酸酶(ALP)活性测定等分析了复合支架材料的生物相容性, 结果表明, MG63细胞在复合支架材料上具有良好的粘附、增殖能力, 并可引起早期的骨分化. 因此, 纳米羟基磷灰石/丝素蛋白复合支架作为骨组织工程的支架材料具有良好的应用前景.     

Effects of Laminin For Osteogenesis in Porous Hydroxyapatite

Summery: As a tooth is composed of hard tissue covering pulp, it may be suitable for tooth regeneration to use porous cylindrical hydroxyapatite (HA) scaffolds with a hollow center. Generally, in vivo examination, bone marrow cell suspension for osteogenesis in cell/HA composite scaffold without subculture is prepared at a density of 1 × 10⁷ cells/ml or higher. In dentistry, stem cells would be obtained from tooth pulp. For dentine formation, a smaller number of stem cells must be used. In this study, a suspension of rat bone marrow cells at 1 × 10⁶ cells/ml of density was prepared to estimate the adhesive effect of laminin. After immersion of HA scaffold in laminin solution, bone marrow cells were seeded in the pores of the HA scaffolds by immersion in the cell suspension for preparing the cell/HA composite scaffolds. The specimens were respectively implanted in the dorsal subcutis of 7-week-old male Fischer 344 rats for 4 weeks for histological examination. Comparing with the results of in vivo examination, alkaline phosphatase activity of bone marrow cells on laminin-coated plate with and without dexamethasone cultured for 2 weeks was measured in vitro. It was considered that laminin contributed to bone formation in pores of a scaffold.     

In-Vitro Biocompatibility Evaluation of Collagen-Hyaluronic Acid/Bioactive Glass Nanocomposite Scaffold

A nano-structured scaffold was designed for bone repair using collagen, hyaluronic acid (HYA) and nano-bioactive glass (NBaG) as its main components. The collagen-HYA/NBaG scaffold was prepared by using a freeze-drying technique and characterized by scanning electron microscopy (SEM). Osteoblastls were seeded on these scaffolds and their proliferation rate, alkaline phosphatase (ALP) activity and ability to form mineralized bone nodules were compared with those osteoblasts grown on cell culture plastic surfaces. The cross-section morphology shows that the collagen-HYA/NBaG scaffold possessed a three-dimensional (3D) interconnected homogenous porous structure. The results obtained from biological assessment show that this scaffold did not negatively affect osteoblasts proliferation rate and improves osteoblasts function as shown by increasing the ALP activity and calcium deposition and formation of mineralized bone nodules. Therefore, the composite scaffolds could provide a favorable environment for initial cell adhesion, maintained cell viability and cell proliferation, and had good in-vitro biocompatibility.     

Porous nano-minerals substituted apatite/chitin/pectin nanocomposites scaffolds for bone tissue engineering

In the current study, a porous 3D scaffold using Gallium-Apatite/chitin/pectin (Ga-HA/C/P) nanocomposites scaffolds (NCS) were fabricated by freeze-drying process with applications in orthopedics (bone tissue engineering). Various NCSs (0%, 30%, 50 and 70%) were prepared and characterized for its chemical structure, crystalline phase, surface texture by using various techniques such as FT-IR, XRD and SEM-EDX, respectively. The analyses of physicochemical properties proved that the formulated scaffolds were highly porous, and mechanically stable with superior density. The nanocomposite scaffolds also presented with increased swelling ability, lower biodegradation rate and higher mechanical strength. Further, biocompatibility and cytotoxicity of Ga-HA/C/P nanocomposite scaffolds were studied using NIH3T3 cells and MG-63 cells revealed no toxicity and cells attached and proliferated on scaffolds. Further implantation of prepared NCS showed mature bone formation through formation of new bone cells and osteoblast differentiation. Also, Ga-HA/C/P nanocomposites scaffolds proved to be more effective than chitin-pectin composite scaffolds. Taking results together it can be inferred that the prepared nanocomposite scaffolds possesses the prerequisites and showed great potential for treating orthopedic applications.     

Fabrication of poly(lactide‐co‐glycolide) scaffold embedded spatially with hydroxyapatite particles on pore walls for bone tissue engineering

Poly(lactide‐co‐glycolide) (PLGA) scaffolds embedded spatially with hydroxyapatite (HA) particles on the pore walls (PLGA/HA‐S) were fabricated by using HA‐coated paraffin spheres as porogens, which were prepared by Pickering emulsion. For comparisons, PLGA scaffolds loaded with same amount of HA particles (2%) in the matrix (PLGA/HA‐M) and pure PLGA scaffolds were prepared by using pure paraffin spheres as porogens. Although the three types of scaffolds had same pore size (450–600 µm) and similar porosity (90%–93%), the PLGA/HA‐S showed the highest compression modulus. The embedment of the HA particles on the pore walls endow the PLGA/HA‐S scaffold with a stronger ability of protein adsorption and mineralization as well as a larger mechanical strength against compression. In vitro culture of rat bone marrow stem cells revealed that cell morphology and proliferation ability were similar on all the scaffolds. However, the alkaline phosphatase activity was significantly improved for the cells cultured on the PLGA/HA‐S scaffolds. Therefore, the method for fabricating scaffolds with spatially embedded nanoparticles provides a new way to obtain the bioactive scaffolds for tissue engineering. Copyright © 2011 John Wiley & Sons, Ltd.     

In vitro blood compatibility and bone mineralization aspects of polymeric scaffold laden with essential oil and metallic particles for bone tissue engineering

Bone tissue engineering scaffolds necessities appropriate physicochemical and mechanical properties to support its renewal. Electrospun scaffolds have been used unequivocally in bone tissue restoration. The main intention of this research is to develop electrospun polyurethane (PU) scaffold decorated with metallic particles and essential oil with advanced properties to make them as a putative candidate. The nanocomposite scaffold exhibited appropriate wettability and suitable fiber diameter compared to the polyurethane scaffold. Interaction of the added constituents with the polyurethane was corroborated through hydrogen bonding formation. Tensile strength of the composites was enhanced compared to the polyurethane scaffold. Thermal analysis depicted the lower weight loss of the composite scaffold than the pristine PU. Blood coagulation was significantly delayed and also the composite surface rendered safe interaction with red blood cells. In vitro toxicity testing using fibroblast cells portrayed the nontoxic behavior of the fabricated material. The above-said advanced properties of the composite scaffold can be warranted for bone tissue engineering application.     

与胶原特异性结合的BMP2模拟肽/PLGA 3D打印复合支架的制备及成骨诱导活性

将胶原绑定结构域(CBD)多肽序列与骨形态发生蛋白2模拟肽(BMP2-MP)序列连接制备具有胶原绑定能力的CBD-BMP2-MP, 再将CBD-BMP2-MP与聚丙交酯-乙交酯/胶原(PLGA/COL)3D打印支架相结合, 以支架表面的胶原成分为媒介, 将CBD-BMP2-MP更有效地固定于骨修复材料上, 达到对其进行改性的目的. 利用扫描电子显微镜(SEM)、 电子万能试验机和接触角测量仪对复合支架表面形貌、 力学强度和亲水性等材料学性能进行评价. 用荧光成像法评测 CBD-BMP2-MP及BMP2-MP与支架材料的结合能力. 在各组支架材料表面接种MC3T3-E1细胞进行体外培养, 采用CCK-8、 鬼笔环肽荧光染色、 茜素红染色及qPCR综合评价细胞在材料表面的黏附、 增殖和成骨分化等细胞行为, 研究CBD-BMP2-MP修饰的3D多孔PLGA/COL复合支架的生物学性能. 研究结果表明, 利用3D打印技术制备的多孔支架具有形貌可控的孔隙结构, 为细胞生长创造更有利的细胞微环境, 支架表面胶原成分的加入提高了支架材料的亲水性, 同时对支架材料本身的力学性能无任何影响, 提高了复合支架本身的生物相容性. 与普通BMP2-MP相比, CBD-BMP2-MP具有更好的胶原绑定能力, 与复合支架的结合更稳定, 提高了PLGA/COL复合支架对BMP2-MP的负载能力. 支架表面负载CBD-BMP2-MP后具有极强的促细胞成骨分化能力. MC3T3-E1细胞表现出更高的钙沉积能力, 并且成骨分化相关基因Runx2, ALP, COL-I及OPN等水平也有了明显提升. 表明CBD-BMP2-MP多孔复合支架具有良好的生物相容性和成骨诱导活性, 在骨组织修复领域具有良好的应用前景.     

Improving mechanical and biological properties of macroporous HA scaffolds through composite coatings

Interconnected porous hydroxyapatite (HA) scaffolds are widely used for bone repair and replacement, owing to their ability to support the adhesion, transfer, proliferation and differentiation of cells. In the present study, the polymer impregnation approach was adopted to produce porous HA scaffolds with three-dimensional (3D) porous structures. These scaffolds have an advantage of highly interconnected porosity (≈85%) but a drawback of poor mechanical strength. Therefore, the as-prepared HA scaffolds were lined with composite polymer coatings in order to improve the mechanical properties and retain its good bioactivity and biocompatibility at the same time. The composite coatings were based on poly(d,l-lactide) (PDLLA) polymer solutions, and contained single component or combination of HA, calcium sulfate (CS) and chondroitin sulfate (ChS) powders. The effects of composite coatings on scaffold porosity, microstructure, mechanical property, in vitro mineralizing behavior, and cell attachment of the resultant scaffolds were investigated. The results showed that the scaffolds with composite coatings resulted in significant improvement in both mechanical and biological properties while retaining the 3D interconnected porous structure. The in vitro mineralizing behaviors were mainly related to the compositions of CS and ChS powders in the composite coatings. Excellent cell attachments were observed on the pure HA scaffold as well as the three types of composite scaffolds. These composite scaffolds with improved mechanical properties and bioactivities are promising bone substitutes in tissue engineering fields.     

Improved efficacy of bio‐mineralization of human mesenchymal stem cells on modified PLLA nanofibers coated with bioactive materials via enhanced expression of integrin α2β1

Current therapeutic interventions in bone defects are mainly focused on finding the best bioactive materials for inducing bone regeneration via activating the related intracellular signaling pathways. Integrins are trans‐membrane receptors that facilitate cell‐extracellular matrix (ECM) interactions and activate signal transduction. To develop a suitable platform for supporting human bone marrow mesenchymal stem cells (hBM‐MSCs) differentiation into bone tissue, electrospun poly L‐lactide (PLLA) nanofiber scaffolds were coated with nano‐hydroxyapatite (PLLA/nHa group), gelatin nanoparticles (PLLA/Gel group), and nHa/Gel nanoparticles (PLLA/nHa/Gel group) and their impacts on cell proliferation, expression of osteoblastic biomarkers, and bone differentiation were examined and compared. MTT data showed that proliferation of hBM‐MSCs on PLLA/nHa/Gel scaffolds was significantly higher than other groups (P < .05). Alkaline phosphatase activity was also more increased in hBM‐MSCs cultured under osteogenic media on PLLA/nHa/Gel scaffolds compared to others. Gene expression evaluation confirmed up‐regulation of integrin α2β1 as well as the osteogenic genes BGLAP, COL1A1, and RUNX2. Following use of integrin α2β1 blocker antibody, the protein level of integrin α2β1 in cells seeded on PLLA/nHa/Gel scaffolds was decreased compared to control, which confirmed that most of the integrin receptors were bound to gelatin molecules on scaffolds and could activate the integrin α2β1/ERK axis. Collectively, PLLA/nHa/Gel scaffold is a suitable platform for hBM‐MSCs adhesion, proliferation, and osteogenic differentiation in less time via activating integrin α2β1/ERK axis, and thus it might be applicable in bone tissue engineering.     

Fabrication and Properties of Porous Collagen/Hyaluronic Composite Scaffolds Containing Nano-Bioactive Glass for Tissue Engineering

In this work, nano-structured scaffolds were designed for tissue engineering using collagen, hyaluronic acid (HA) and nano-bioactive glass (NBAG) as their main components. The scaffold was prepared via freeze-drying method and the properties including morphology, porosity, compressive strength, swelling ratio and cytotoxicity in-vitro, were also evaluated. The composite scaffolds showed well interconnected macropores with the pore size of ranging from 100 to 500 μm. The porosity percent and swelling ability were decreased with the introduction of NBAG into the collagen/HA hydrogel; however, the compressive strength was enhanced. The cytotoxicity in-vitro study shows that the collagen-HA/NBAG scaffolds have good biocompatibility with improving effect on fibroblastic cells growth. It could be concluded that this scaffold fulfills the main requirements to be considered as a bone substitute.     

Reinforcement of electrospun polycaprolacton scaffold using KIT-6 to improve mechanical and biological performance

Electrospinning has been extensively accepted as one of most important techniques for fabrication of scaffolds for bone tissue engineering. Polycaprolactone is one of the most applied electro-spinned scaffolds. Since low mechanical strength of polycaprolactone scaffold leads to the limitation of its applications, composition of polycaprolactone with ceramic particles is of great interest. Several studies have been conducted on fabrication and characterization of polycaprolactone nanocomposite scaffolds, but none of these researches has used mesoporous silica particles (KIT-6). In this project, a high-strength and bioactive nanocomposite scaffold has been developed which consists of polycaprolactone and mesoporous silica particles. Results showed that increase of KIT-6 particles percentages up to 5% leads to the enhancement of tensile strength of scaffold from 1.8 ± 0.2 to 2.9 ± 1.0 MPa. Although wettability of scaffolds in presence of particles was totally lower than pure PCL scaffold, but increase of particles percentages led to enhancement of wettability and water absorption of scaffolds. On the other hand presence of KIT-6 particles increased specific surface area and also bioactivity of scaffold was increased by enhancement of ion exchange between surface and simulated body fluid. Finally it was concluded that PCL-KIT-6 scaffolds are a suitable candidate for application in tissue engineering.     

Synthesis and characterization of a nano‐hydroxyapatite/chitosan/polyethylene glycol nanocomposite for bone tissue engineering

A novel nanocomposite involving nano‐hydroxyapatite/chitosan/polyethylene glycol (n‐HAP/CS/PEG) has been successfully synthesized via co‐precipitation approach at room temperature. The purpose to synthesize such nanocomposite is to search for an ideal analogue which may mimick the composition of natural bone for bone tissue engineering with respect to suitable biocompatibility, cytotoxicity and mechanical properties. The FTIR spectra of n‐HAP/CS and n‐HAP/CS/PEG scaffolds indicated significant intermolecular interaction between the various components of both the nanocomposites. The results of XRD, TEM and TGA/DTA suggested that the crystallinity and thermal stability of the n‐HAP/CS/PEG scaffold have decreased and increased respectively, relative to n‐HAP/CS scaffold. The comparison of SEM images of both the scaffolds indicated that the incorporation of PEG influenced the surface morphology while a better in‐vitro bioactivity has been observed in n‐HAP/CS/PEG than in n‐HAP/CS based on SBF study, referring a greater possibility for making direct bond to living bone if implanted. Furthermore, MTT assay revealed superior non‐toxic nature of n‐HAP/CS/PEG to murine fibroblast L929 cells as compared to n‐HAP/CS. The comparative swelling studies of n‐HAP/CS/PEG and n‐HAP/CS scaffolds revealed a better swelling rate for n‐HAP/CS/PEG. Also n‐HAP/CS/PEG showed higher mechanical strength relative to n‐HAP/CS supportive of bone tissue ingrowths. Copyright © 2014 John Wiley & Sons, Ltd.     

Poly(l‐Lactic Acid)/Gelatin Fibrous Scaffold Loaded with Simvastatin/Beta‐Cyclodextrin‐Modified Hydroxyapatite Inclusion Complex for Bone Tissue Regeneration

Recently, the application of nanostructured materials in the field of tissue engineering has garnered attention to mediate treatment and regeneration of bone defects. In this study, poly(l ‐lactic acid) (PLLA)/gelatin (PG) fibrous scaffolds are fabricated and β‐cyclodextrin (βCD) grafted nano‐hydroxyapatite (HAp) is coated onto the fibrous scaffold surface via an interaction between βCD and adamantane. Simvastatin (SIM), which is known to promote osteoblast viability and differentiation, is loaded into the remaining βCD. The specimen morphologies are characterized by scanning electron microscopy. The release profile of SIM from the drug loaded scaffold is also evaluated. In vitro proliferation and osteogenic differentiation of human adipose derived stem cells on SIM/HAp coated PG composite scaffolds is characterized by alkaline phosphatase (ALP) activity, mineralization (Alizarin Red S staining), and real time Polymerase chain reaction (PCR). The scaffolds are then implanted into rabbit calvarial defects and analyzed by microcomputed tomography for bone formation after four and eight weeks. These results demonstrate that SIM loaded PLLA/gelatin/HAp‐(βCD) scaffolds promote significantly higher ALP activity, mineralization, osteogenic gene expression, and bone regeneration than control scaffolds. This suggests the potential application of this material toward bone tissue engineering.

     

新型介孔羟基磷灰石/壳聚糖-万古霉素药物释放系统复合材料的制备及体外抗菌和成骨能力

采用冷冻干燥法合成了介孔羟基磷灰石(HA)/壳聚糖(CS)-万古霉素(VCM)药物释放系统复合材料, 利用SEM, XRD和FTIR等方法对材料进行了表征. 结果证实CS与HA混合复合材料具有良好的孔径和孔隙率, 万古霉素吸附于复合材料的表面和内部. 细胞毒性实验[噻唑蓝(MTT)比色法]结果表明, 材料可以促进成骨细胞增殖且具有良好的细胞相容性. 体外抑菌实验结果证实此材料可长时间抑制耐甲氧西林金葡菌(MRSA)的生长, 具有良好的抑菌和杀菌能力. 细胞黏附实验结果表明, 成骨细胞附着于材料表面增殖并通过孔道延伸. 实时聚合酶链式反应(RT-PCR)实验结果表明, 在成骨相关标志产物胶原蛋白-1(COL-1)及骨形态发生蛋白-2(BMP-2)基因上均有较高的表达, 表明材料在体外可以促进成骨细胞生长, 具有良好的成骨能力.     

Controllable synthesis and characterization of porous polyvinyl alcohol/hydroxyapatite nanocomposite scaffolds via an in situ colloidal technique

During the last decades, there have been several attempts to combine bioactive materials with biocompatible and biodegradable polymers to create nanocomposite scaffolds with excellent biocompatibility, bioactivity, biodegradability and mechanical properties. In this research, the nanocomposite scaffolds with compositions based on PVA and HAp nanoparticles were successfully prepared using colloidal HAp nanoparticles combined with freeze-drying technique for tissue engineering applications. In addition, the effect of the pH value of the reactive solution and different percentages of PVA and HAp on the synthesis of PVA/HAp nanocomposites were investigated. The SEM observations revealed that the prepared scaffolds were porous with three dimensional microstructures, and in vitro experiments with osteoblast cells indicated an appropriate penetration of the cells into the scaffold's pores, and also the continuous increase in cell aggregation on the scaffolds with increase in the incubation time demonstrated the ability of the scaffolds to support cell growth. According to the obtained results, the nanocomposite scaffolds could be considered as highly bioactive and potential bone tissue engineering implants.     

Development of nano-tricalcium phosphate/polycaprolactone/platelet-rich plasma biocomposite for bone defect regeneration

Nano-tricalcium phosphate (n-TCP) is an osteoconductive substance which, like polycaprolactone (PCL), has been used for clinical purposes for many years; It has now been licensed for a range of products for clinical and medication distribution. This research aimed to examine the effects of platelet-rich plasma on mesenchymal stem cell proliferation and osteogenic differentiation. Thus, we decided to examine the in vitro and in vivo actions of PRP-treated porous biocomposite scaffolds based on nano-tricalcium phosphate- polycaprolactone (n-TCP-PCL/PRP). The prepared samples were described utilizing FTIR, XRD, and SEM. MTT has measured the cytotoxicity of the biocomposite scaffolds. After two weeks of cell seeding, Alizarin red staining confirmed bone mineral formation by MSCs cells. Moreover, from day 4 to day 7, n-TCP-PCL/PRP biocomposite scaffold improved the expresses of bone marker genes. Platelet-rich plasma (PRP) in conjunction with nano-tricalcium phosphate- polycaprolactone (n-TCP-PCL) biocomposite scaffold is beneficial for the regeneration and stability of the freshly developed bone tissue.