可注射nHA/PU复合多孔骨修复支架制备及表征

兼具良好孔隙率和原位任意塑形固化的可注射复合多孔骨修复材料在临床不规则骨缺损的治疗方面显示出巨大的优势。本研究通过优化双组分设计,以水为发泡剂制备可注射纳米羟基磷灰石/聚氨酯（nHA/PU）复合多孔支架。利用扫描电子显微镜（SEM）、傅里叶变换红外光谱（FTIR）、X射线衍射（XRD）、力学测试及Gillmore针测试等手段对制备的支架进行结构形貌、化学组成、力学性能和固化时间表征。结果表明,本研究制备的可注射nHA/PU复合多孔支架孔隙率高、孔隙贯通性好,孔径分布在100~700μm,适宜细胞和组织向孔内生长;添加20% nHA显著提高了PU支架的力学强度,但降低了支架的孔隙率;可注射支架在8h固化,适宜临床操作。本研究制备的可注射nHA/PU复合多孔支架在不规则骨缺损修复领域具有较大的应用潜力。     

In Vitro Degradation of PLCL/nHA Biodegradable Scaffolds

We investigated the effect of bioactive nanoparticles on the in-vitro degradation of PLCL and PLCL/nHA composite scaffolds. The concentration of nanohydroxyapatite significantly affected the degradation rate. An increase in the crystallinity of the amorphous portion of the polymer was observed. This increased crystallinity was more pronounced in the pure PLCL samples than in those with more nHA. During the degradation process, we observed the appearance of multiple micropores on the scaffold walls as the hydrolysis process progressed and, by the sixth week, the remains of the degradation products were visible on the pore walls.     

Evaluation of poly(L-lactide) and chitosan composite scaffolds for cartilage tissue regeneration

The present study delineates the development of chitosan and poly(L-lactide) (PLLA) scaffolds cross-linked using a mixture of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), n-hydroxysuccinimide (NHS), and chondroitin sulfate (CS) for cartilage tissue engineering applications. Chitosan and PLLA were varied in concentration for developing scaffolds and prepared by freeze-drying method. The various scaffolds were studied using scanning electron microscopy (SEM), porosity by mercury intrusion porosimeter, and the molecular interactions among polymers using Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) studies. Differential scanning calorimetry was used to predict the thermal properties of the scaffolds. The mechanical properties of the scaffolds were studied using static mechanical tester. The ability of the scaffolds to support chondrocyte proliferation was also studied. The microscopy suggests that the pore size of the scaffolds varied with the composition in the range of 38–172 μm and the porosities in the range of 73–93%. The XRD and the FTIR studies suggested that an alternation in the composition of the scaffolds altered the molecular interactions among the scaffold components. An increase in the chitosan content enhanced the swelling property. The degradation of the scaffolds was least when the proportion of chitosan and PLLA was in the ratio of 70:30. The in vitro cell proliferation study suggested that the developed scaffolds were able to support chondrogenesis, the glycosaminoglycan (GAG) content of the mature chondrocyte was 40 μg/ml and the viability was approximately 90%. Hence, the so designed scaffolds may be tried for cartilage tissue engineering applications.     

Preparation and characterization of plla composite scaffolds by ScCO2‐induced phase separation

Composite Scaffolds have received much attention in the tissue engineering, and how to choose the materials has become the research focus in this field. Supercritical CO₂ (ScCO₂)‐induced phase separation process was employed to prepare porous poly‐L ‐lactide (PLLA) composite scaffolds. An experiment system was set up for the purpose of investigating the effects of such parameters as the mass ratios of PLLA to polyethylene glycol (PEG) and PLLA to β‐TCP on porosity and compressive strength of composite scaffolds. The obtained composite scaffolds were characterized in many ways. Scanning electron microscopy was used to examine the morphology and pore size; porosity was analyzed by pycnometer; and the compressive strength was recorded by texture analyzer. The results indicated that the porosity was increased with the addition of PEG, and the highest porosity of PLLA/PEG composite scaffolds was 92% with the mass ratio of PLLA to PEG of 1:0.05; the compressive strength was increased with the addition of β‐TCP, and the highest compressive strength of PLLA/β‐TCP composite scaffolds was 1.76 MPa with the mass ratio of PLLA to β‐TCP of 1:0.1. POLYM. COMPOS., 2012. © 2012 Society of Plastics Engineers     

Effect of Poly(L-lactic acid) on Hydrolytic Degradation of Poly(glycolic acid)

The in-vitro degradation behavior of poly(glycolic acid) (PGA) rods and the composite rods containing poly(L-lactic acid) (PLLA) were investigated via mass loss, pH value change, scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). Since the degradation rate of PLLA is lower than that of PGA, PLLA/PGA composite rods exhibit a slower degradation rate in comparison with PGA. This finding indicated that it was possible to control the degradation rate of the composites by changing their composition. This result indicates that this kind of composite biomaterial may be applicable to devices for the need of prolonged degradation.     

PLGA/nHA hybrid nanofiber scaffold as a nanocargo carrier of insulin for accelerating bone tissue regeneration

The development of tissue engineering in the field of orthopedic surgery is booming. Two fields of research in particular have emerged: approaches for tailoring the surface properties of implantable materials with osteoinductive factors as well as evaluation of the response of osteogenic cells to these fabricated implanted materials (hybrid material). In the present study, we chemically grafted insulin onto the surface of hydroxyapatite nanorods (nHA). The insulin-grafted nHAs (nHA-I) were dispersed into poly(lactide-co-glycolide) (PLGA) polymer solution, which was electrospun to prepare PLGA/nHA-I composite nanofiber scaffolds. The morphology of the electrospun nanofiber scaffolds was assessed by field emission scanning electron microscopy (FESEM). After extensive characterization of the PLGA/nHA-I and PLGA/nHA composite nanofiber scaffolds by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction spectroscopy (XRD), X-ray photoelectron spectroscopy (XPS), energy-dispersive X-ray spectrometry (EDS), and transmission electron microscopy (TEM), the PLGA/nHA-I and PLGA/nHA (used as control) composite nanofiber scaffolds were subjected to cell studies. The results obtained from cell adhesion, alizarin red staining, and Von Kossa assay suggested that the PLGA/nHA-I composite nanofiber scaffold has enhanced osteoblastic cell growth, as more cells were proliferated and differentiated. The fact that insulin enhanced osteoblastic cell proliferation will open new possibilities for the development of artificial scaffolds for bone tissue regeneration.     

I‐Optimal design of poly(lactic‐co‐glycolic) acid/hydroxyapatite three‐dimensional scaffolds produced by thermally induced phase separation

In bone tissue engineering, three‐dimensional (3D) scaffolds are often designed to have adequate modulus while taking into consideration the requirement for a highly porous network for cell seeding and tissue growth. This article presents the design optimization of 3D scaffolds made of poly(lactic‐co‐glycolic) acid (PLGA) and nanohydroxyapatite (nHA), produced by thermally induced phase separation (TIPS). Slow cooling at a rate of 1°C/min enabled a uniform temperature and produced porous scaffolds with a relatively uniform pore size. An I‐optimal design of experiments (DoE) with 18 experimental runs was used to relate four responses (scaffold thickness, density, porosity, and modulus) to three experimental factors, namely the TIPS temperature (?20, ?10, and 0°C), PLGA concentration (7%, 10%, and 13% w/v), and nHA content (0%, 15%, and 30% w/w). The response surface analysis using JMP® software predicted a temperature of ?18.3°C, a PLGA concentration of 10.3% w/v, and a nHA content of 30% w/w to achieve a thickness of 3 mm, a porosity of 83%, and a modulus of ~4 MPa. The set of validation scaffolds prepared using the predicted factor levels had a thickness of 3.05 ± 0.37 mm, a porosity of 86.8 ± 0.9%, and a modulus of 3.57 ± 2.28 MPa. POLYM. ENG. SCI., 59:1146–1157 2019. © 2019 Society of Plastics Engineers     

Preparation and characterization of aliphatic polyurethane and hydroxyapatite composite scaffold

Novel porous composite scaffolds for tissue engineering were prepared from aliphatic biodegradable polyurethane (PU) elastomer and hydroxyapatite (HA). It was found that the aliphatic PU was possible to load up to 50 wt % HA. The morphology and properties of the scaffolds were characterized by scanning electron microscope, X‐ray diffraction, infrared absorption spectra, mechanical testing, dynamic mechanical analysis, and in vitro degradation measurement. The results indicated that the HA/PU scaffolds had an interconnected porous structure with a pore size mainly ranging from 300 to 900 μm, and 50–200 μm micropores existed on the pores' walls. The average pore size of macropores and micropores are 510 and 100 μm, respectively. The compressive strength of the composite scaffolds showed higher enhancement with increasing HA content. In addition, the polymer matrix was completely composed of aliphatic component and exhibited progressive mass loss in vitro degradation, and the degradation rate depended on the HA content in PU matrix. The porous HA/PU composite may have a good prospect to be used as scaffold for tissue engineering. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Fabrication of macroporous chitosan/poly(l-lactide) hybrid scaffolds by sodium acetate particulate-leaching method

For tissue engineering applications, 3-D macroporous chitosan/poly(l-lactide) (PLLA) scaffolds were prepared by the particulate-leaching method using sodium acetate as the porogen in an acidic water/dioxane solution. The stability and dispersity of chitosan on the chitosan/PLLA hybrid scaffolds were determined by measuring water contact angles, establishing crystallinity using X-ray diffraction, and using eosin staining to observe the chitosan under a light microscope. The porous structure of the particulate-leached chitosan/PLLA scaffolds was investigated in terms of pore morphology, interconnectivity, etc. by using scanning electron microscopy. Chitosan/PLLA scaffolds produced by particulate-leaching showed a highly porous structure and improved stability and dispersity of chitosan as compared to pure PLLA and chitosan-coated PLLA scaffolds. The highly porous structure that resulted from a high concentration of chitosan improved the efficiency of cell adhesion after culturing cells for 4?h. After 48?h, the cultured cells showed increased cell proliferation on the hybrid scaffolds. Thus, particulate-leached chitosan/PLLA scaffolds can be applied to tissue engineering of various types, including the industrial membrane field.     

Preparation of Pure PLLA,Pure Chitosan,and PLLA/Chitosan Blend Porous Tissue Engineering Scaffolds by Thermally Induced Phase Separation Method and Evaluation of the Corresponding Mechanical and Biological Properties

The aim of this work was to prepare the scaffolds of pure poly (L-lactic acid) 3% (w/v), pure chitosan 3% (w/v), and PLLA/chitosan blend (1:5) 3% (w/v) using TIPS method and investigate their properties and application in tissue engineering. An in vitro degradation study of scaffolds showed the addition of chitosan to PLLA not only increased its degradation rate, but also slowed down its pH value reduction. Addition of chitosan to PLLA increased hydrophilicity, porosity, compressive properties, and cell viability of the scaffolds. The results indicate that among all scaffolds, the most appropriate candidate for tissue engineering is PLLA/chitosan blend.     

Preparation of thermoplastic polyurethane/graphene oxide composite scaffolds by thermally induced phase separation

In this study, biomedical thermoplastic polyurethane/graphene oxide (TPU/GO) composite scaffolds were successfully prepared using the thermally induced phase separation (TIPS) technique. The microstructure, morphology, and thermal and mechanical properties of the scaffolds were characterized by Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), and compression tests. Furthermore, NIH 3T3 fibroblast cell viability on the porous scaffolds was investigated via live/dead fluorescent staining and SEM observation. FTIR and Raman results verified the presence of GO in the composites. SEM images showed that the average pore diameter of the composite scaffolds decreased as the amount of GO increased. Additionally, the surface of the specimens became rougher due to the embedded GO. The compressive modulus of composite specimens was increased by nearly 200% and 300% with the addition of 5% and 10% GO, respectively, as compared with pristine TPU. 3T3 fibroblast culture results showed that GO had no apparent cytotoxicity. However, high loading levels of GO may delay cell proliferation on the specimens. POLYM. COMPOS., 35:1408–1417, 2014. © 2013 Society of Plastics Engineers     

Protein expression by bone mesenchymal stem cells cultivated in PLLA scaffolds with different pore geometry

Abstract

The effects of poly(L,L-lactide) (PLLA) scaffold with axial and isotropic structure were investigated on functional activity of rabbit bone mesenchymal stem cells (BMSCs). PLLA scaffolds were processed by freeze-dry technique at different temperatures of the scaffold frost – ?196?°C, ?25?°C and 0?°C. Scaffolds with different pore sizes were obtained by adding 5 or 10% of water phase. Scaffolds were modified by collagen type I solution. The pore sizes of polymer scaffolds were ranging from 5 to 150?µm. More protein secretion was observed in the surface-modified scaffolds than in the unmodified after 2 weeks of cultivation in vitro.     

Injection‐molded porous hydroxyapatite/polyamide‐66 scaffold for bone repair and investigations on the experimental conditions

Hydroxyapatite/polyamide‐66 (HA/PA66) composite scaffolds were prepared using injection‐molding technique and also analyzed by means of scanning electron microscopy, X‐ray diffraction, differential scanning calorimetry, Fourier transform infrared spectroscopy, and mechanical testing. Compared with common methods to fabricate scaffolds, this process can fabricate composite scaffolds in a rapid and convenient manner by adjusting the experimental conditions of foaming agent and shot size. The interactions between PA66 and HA particles affect the crystallization behavior of PA66 and the pore structure of scaffolds. HA particles can increase the stiffness of composite scaffolds accompanied by the reduction of impact strength, pore size and porosity. The obtained 40 wt% HA/PA66 composite scaffolds with a pore size ranging from 100–500 μm and a porosity more than 65% can simultaneously meet the requirements of porous structure and mechanical performance. POLYM. ENG. SCI., 54:1003–1012, 2014. © 2013 Society of Plastics Engineers     

Electrospun poly(L‐lactic acid)/hydroxyapatite composite fibrous scaffolds for bone tissue engineering

Poly(L ‐lactic acid) (PLLA) is one of the most studied synthetic biodegradable polymeric materials as a bone graft substitute. Taking into account the osteoconductive property of hydroxyapatite (HAp), we prepared fibrous matrices of PLLA without and with HAp particles in amounts of 0.25 or 0.50% (w/v, based on the volume of the base 15% w/v PLLA solution in 70:30 v/v dichloromethane/tetrahydrofuran). These fibrous matrices were assessed for their potential as substrates for bone cell culture. The presence of HAp in the composite fibre mats was confirmed using energy dispersive X‐ray spectroscopy mapping. The average diameters of both neat PLLA and PLLA/HAp fibres, as determined using scanning electron microscopy, ranged between 2.3 and 3.5 µm, with the average spacing between adjacent fibres ranging between 5.7 and 8.5 µm. The porosity of these fibrous membranes was high (ca 97–98%). A direct cytotoxicity evaluation with L929 mouse fibroblasts indicated that the neat PLLA fibre mats released no substance at a level that was toxic to the cells. The presence of HAp particles at 0.50% w/v in the PLLA fibrous scaffolds not only promoted the attachment and the proliferation of MC3T3‐E1 mouse pre‐osteoblastic cells, but also increased the expression of osteocalcin mRNA and the extent of mineralization after the cells had been cultured on the scaffolds for 14 and 21 days, respectively. The results obtained suggested that the PLLA/HAp fibre mats could be materials of choice for bone tissue engineering. Copyright © 2009 Society of Chemical Industry     

Physical,morphological, and biological studies on PLA/nHA composite nanofibrous webs containing Equisetum arvense herbal extract for bone tissue engineering

A series of herbal extract incorporated into poly(lactic acid) (PLA) composite nanofibrous scaffolds were successfully prepared by using electrospinning technique. Equisetum arvense extract (EE) and nanohydroxyapatite (nHA) in different quantities were loaded into PLA solution to fabricate composite nanofibrous webs under various electrospinning conditions. Uniform nanofibers were obtained with an average diameter of 157 ± 47 nm in the case of those containing the herbal extract. Characterization of the webs was carried out by means of Fourier transform infrared (FTIR) spectroscopy, field emission‐scanning electron microscopy (FESEM), energy‐dispersive X‐ray spectroscopy (EDX), and differential scanning calorimetry (DSC) techniques. Mechanical properties, porosity, and contact angle of the prepared webs were also determined. Releasing behavior was investigated in phosphate buffer solution (pH 7.2) medium. Moreover, cell studies and osteogenic capacity were assessed in vitro using human adipose tissue‐derived mesenchymal stem cell (AT‐MSC). Evaluations of cell attachment, spreading, and proliferation of AT‐MSC were done by SEM observation and thiazolyl blue (MTT) assay. Osteogenic differentiation capability of AT‐MSC on the nanofibrous webs was analyzed by alkaline phosphatase activity and calcium content assay. It was found that with the addition of nHA and EE to PLA nanofibrous webs, their surface hydrophobicity was reduced while the tensile strength and Young's modulus were increased satisfactorily. Regarding the samples containing EE and nHA, cellular adhesion was observed with flattened normal morphology. Osteogenic differentiation of AT‐MSC on PLA/nHA/EE webs showed the highest mineralization capacity after 3 weeks which, was about 1.8 and 3 times higher than that of PLA/nHA and tissue culture polystyrene as control, respectively. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45343.     

Microstructure and mechanical properties of poly(L‐lactide) scaffolds fabricated by gelatin particle leaching method

Biodegradable poly(L ‐lactide) (PLLA) scaffolds with well‐controlled interconnected irregular pores were fabricated by a porogen leaching technique using gelatin particles as the porogen. The gelatin particles (280–450 μm) were bonded together through a treatment in a saturated water vapor condition at 70°C to form a 3‐dimensional assembly in a mold. PLLA was dissolved in dioxane and was cast onto the gelatin assembly. The mixtures were then freeze‐dried or dried at room temperature, followed by removal of the gelatin particles to yield the porous scaffolds. The microstructure of the scaffolds was characterized by scanning electron microscopy with respect to the pore shape, interpore connectivity, and pore wall morphology. Compression measurements revealed that scaffolds fabricated by freeze‐drying exhibited better mechanical performance than those by room temperature dying. Along with the increase of the polymer concentration, the porosity of the scaffolds decreased whereas the compressive modulus increased. When the scaffolds were in a hydrated state, the compressive modulus decreased dramatically. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 98: 1373–1379, 2005     

Fabrication and evaluation of nanofibrous polyhydroxybutyrate valerate scaffolds containing hydroxyapatite particles for bone tissue engineering

Tissue engineering is a new approach for regeneration of damaged tissues. The current clinical methods such as autograft and allograft transplantation are not effective for repairing bone damages, mainly due to the limited available sources and the donor-site side effects. In this research, the nanocomposite poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)/nano hydroxyapatite (nHA) scaffolds with different nHA ratios for bone regeneration were utilized. The diameter and porosity of scaffolds were approximately 200?nm and 74%, respectively. The degradability test of the scaffolds suggests a low degradation rate with total degradation of 30% after 3 months. Cytotoxicity result showed that cultured osteoblast cells (MC3T3) on nanocomposite scaffolds had superiority in terms of higher proliferation and attachment in comparison with PHBV scaffold. The protein expression of alkaline phosphatase illustrated that nanofibrous scaffold containing hydroxyapatite had the highest alkaline phosphatase activities as a result of better proliferation. These results recommend that PHBV/nHA scaffolds are suitable candidates for bone tissue engineering.     

Enhanced degradation of poly(L‐lactide) containing arginine,tryptophan and lysine

Addition of arginine, tryptophan and lysine to poly(L ‐lactide) (PLLA) has resulted in a new series of enhanced degradation biomaterials. Degradation was performed at 70°C and degradation behavior was studied using water uptake, mass loss, pH value change, scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and tensile properties. The results indicated that PLLA containing amino acids degraded more quickly than pure PLLA. In PLLA containing arginine, lysine, or tryptophan, crystallinity of the latter decreased after 8 days degradation while crystallinities of the other two composites increased. Destroying crystallites assists with deeper degradation and in vivo assimilation. Initial tensile strengths of all undegraded bars were almost the same. Furthermore, pH value changes indicated that additive amino acids could neutralize acidic degradation products, which may be a solution to the bacteria‐free inflammation induced by the acidic products. The results indicate that the composite biomaterials may be useful for future clinical application. POLYM. COMPOS., 2009. © 2008 Society of Plastics Engineers     

Improvement of antifouling performance of poly(l‐lactic acid) membranes through incorporating polyaniline nanoparticles

Poly(l ‐lactic acid) (PLLA) composite membranes were fabricated by nonsolvent induced phase separation method using polyaniline (PANI) as an additive. Membrane structure was characterized by attenuated total reflectance Fourier transform‐infrared spectroscopy, X‐ray diffraction, scanning electron microscopy, porosity, and pore size analysis. Membrane performance was assessed by goniometer, pure water flux, molecular weight cut‐off, static adsorption and dynamic filtration. The incorporation of PANI significantly improved the hydrophilicity and permeability of PLLA composite membrane, and eventually enhanced the antifouling performance of composite membrane compared with pure PLLA membrane. It was demonstrated that PLLA composite membrane with 1 wt % PANI had better separation and antifouling performance compared with other composite membranes. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44452.     

Nanohydroxyapatite-coated hydroxyethyl cellulose/poly (vinyl) alcohol electrospun scaffolds and their cellular response

The present study focused on the preparation of nanohydroxyapatite (nHA)-coated hydroxyethyl cellulose/polyvinyl alcohol (HEC/PVA) nanofibrous scaffolds for bone tissue engineering application. The electrospun HEC/PVA scaffolds were mineralized via alternate soaking process. FESEM revealed that the nHA was formed uniformly over the nanofibers. The nHA mineralization enhanced the tensile strength and reduced the elongation at breakage of scaffolds. The wettability of the nanofibrous scaffolds was significantly improved. The in vitro biocompatibility of scaffolds was evaluated with human osteosarcoma cells. nHA-coated scaffolds had a favorable effect on the proliferation and differentiation of osteosarcoma cell and could be a potential candidate for bone regeneration.