Rapid treatment of full‐thickness skin loss using ovine tendon collagen type I scaffold with skin cells

The full‐thickness skin wound is a common skin complication affecting millions of people worldwide. Delayed treatment of this condition causes the loss of skin function and integrity that could lead to the development of chronic wounds or even death. This study was aimed to develop a rapid wound treatment modality using ovine tendon collagen type I (OTC‐I) bio‐scaffold with or without noncultured skin cells. Genipin (GNP) and carbodiimide (EDC) were used to cross‐link OTC‐I scaffold to improve the mechanical strength of the bio‐scaffold. The physicochemical, biomechanical, biodegradation, biocompatibility, and immunogenicity properties of OTC‐I scaffolds were investigated. The efficacy of this treatment approach was evaluated in an in vivo skin wound model. The results demonstrated that GNP cross‐linked OTC‐I scaffold (OTC‐I_GNP) had better physicochemical and mechanical properties compared with EDC cross‐linked OTC‐I scaffold (OTC‐I_EDC) and noncross‐link OTC‐I scaffold (OTC‐I_NC). OTC‐I_GNP and OTC‐I_NC demonstrated no toxic effect on cells as it promoted higher cell attachment and proliferation of both primary human epidermal keratinocytes and human dermal fibroblasts compared with OTC‐I_EDC. Both OTC‐I_GNP and OTC‐I_NC exhibited spontaneous formation of bilayer structure in vitro. Immunogenic evaluation of OTC‐I scaffolds, in vitro and in vivo, revealed no sign of immune response. Finally, implantation of OTC‐I_NC and OTC‐I_GNP scaffolds with noncultured skin cells demonstrated enhanced healing with superior skin maturity and microstructure features, resembling native skin in contrast to other treatment (without noncultured skin cells) and control group. The findings of this study, therefore, suggested that both OTC‐I scaffolds with noncultured skin cells could be promising for the rapid treatment of full‐thickness skin wound.     

海洋生物医用材料的临床应用和安全性评价

目的： 对海洋生物医用材料在医疗领域的应用情况和海洋生物材料来源医疗器械的安全性评价趋势进行分析，为推进该材料的临床转化提供参考。方法： 归纳海洋生物医用材料的分类和应用，介绍该材料的安全性评价的程序要点，探讨其安全性评价中面临的挑战。结果与结论： 常用的海洋生物医用材料主要为多糖和蛋白质，在创伤修复和组织工程领域应用广泛。海洋生物医用材料具有生物活性和良好的生物相容性，对此类材料的安全性评价应根据材料特性和预期用途，科学制定评价程序和选择检验方法。     

Effect of topology of poly(L-lactide-co-ε-caprolactone) scaffolds on the response of cultured human umbilical cord Wharton’s jelly-derived mesenchymal stem cells and neuroblastoma cell lines

In this study, for the first time, a biodegradable poly(L-lactide-co-ε-caprolactone), PLC 67:33 copolymer was developed for use as temporary scaffolds in reconstructive nerve surgery. The effect of the surface topology and pore architecture were studied on the biocompatibility for supporting the growth of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs) and human neuroblastoma cells (hNBCs) as cell models. Porous PLC membranes were prepared by electrospinning and phase immersion precipitation with particulate leaching and nonporous PLC membranes were prepared by solvent casting. From the results, the porous PLC membranes can support hWJ-MSCs and hNBCs cells better than the nonporous PLC membrane, and the interconnected pore scaffold prepared by electrospinning exhibited a more significant supporting attachment of the cells than the open pore and nonporous membranes. We can consider that these electrospun PLC membranes with 3-D interconnecting fiber networks and a high porosity warrant a potential use as nerve guides in reconstructive nerve surgery.     

皮下种植复方羟磷灰石钛尖根管充填材料的组织学反应

目的：观察复方羟磷灰石钛尖的组织相容性。方法：将填有实验材料或ｏｘｐａｒａ牙胶尖的硅胶管埋植于猴皮下组织内，３个月后，镜下观察组织学反应。结果：实验组不论充填程度如何，均无明显炎症或反应甚微，而对照组的反应较大。结论：复方羟磷灰石钛尖是一种良好的生物相容性根管充填料。     

Biocompatibility of resin-based materials used as pulp-capping agents

AIM: To evaluate and compare the response of pulps of rats capped with resin-modified glass-ionomer cement (RMGIC) or self-etching adhesive system. METHODOLOGY: Class I cavities were prepared on the occlusal surface of 54 maxillary first molars of 27 rats. Pulp exposure was performed on the cavity floor. The following resin-based materials were applied as pulp-capping agents: G1, Clearfil Liner Bond 2V (CLB 2V; Kuraray Co., Japan); G2, Vitrebond (VIT; 3M/ESPE, USA). In group 3 (control group), a calcium hydroxide/saline paste (CH; Labsynth, Brazil) was used. The cavities were restored with amalgam. After 7, 30 and 60 days, the animals were sacrificed and the jaws were processed for microscopic evaluation. RESULTS: Despite the inflammatory response caused by the experimental and the control materials at 7 days, pulpal healing associated with calcified barrier formation was observed at 60 days following the pulp therapy. Both resin-based materials promoted a large zone of cell-rich fibrodentine matrix deposition on the pulp horn related to the pulp exposure site, which was larger to VIT than to CLB 2V specimens. Tertiary dentine underneath the fibrodentine matrix was deposited by a layer of elongated pulpal cells. The remaining pulpal tissue exhibited normal histological characteristics. In the control group, healing and dentine-bridge formation was observed at 30 days. Pulpal breakdown occurred only when bacterial infection occurred. CONCLUSION: Both experimental pulp-capping agents allowed pulpal healing characterized by cell-rich fibrodentine and tertiary dentine deposition as well as calcified barrier formation.     

Effect of ProRoot MTA mixed with chlorhexidine on apoptosis and cell cycle of fibroblasts and macrophages in vitro

AIM: To compare the percentage of apoptotic cells and the cell cycle profile of fibroblasts and macrophages exposed to either ProRoot mineral trioxide aggregate (MTA) mixed with chlorhexidine (CHX), or exposed to ProRoot MTA mixed with sterile water. METHODOLOGY: Mouse gingival fibroblasts or mouse macrophages were seeded in six-well plates and allowed to attach overnight. Freshly mixed or set (allowed to dry for 24 h) specimens of tooth-coloured (white) ProRoot MTA were prepared with 0.12% CHX gluconate (MTA/CHX) or with sterile water (MTA/H2O). The cells were exposed for 24 h to the MTA specimens, which were placed over permeable membrane inserts to avoid direct contact with the cells. Untreated cells served as controls. Propidium iodide staining followed by flow cytometry was used to evaluate the effects of ProRoot MTA on cell apoptosis and cell cycle. Statistical analyses were performed by one-way anova followed by post-hoc tests with the use of the SigmaStat 2.0 software, and significance was determined at P < or = 0.05. RESULTS: MTA specimens containing CHX induced apoptosis of macrophages and fibroblasts (P < 0.05). In contrast, no change in the proportion of apoptotic cells was observed when sterile water was used to prepare the specimens (P > 0.05). Cell cycle analysis showed that exposure to MTA/CHX decreased the percentage of fibroblasts and macrophages in S phase (DNA synthesis) as compared with exposure to MTA/H2O (P < 0.05). CONCLUSION: This in vitro study demonstrated that the substitution of CHX for sterile water in MTA increases its cytotoxicity. This suggests that the potentially beneficial antimicrobial effect of CHX may be accompanied by an increase in the cytotoxicity of the resulting MTA-based material.     

粗化处理对新型骨植入钛合金的生物相容性影响

目的研究国内新研制的骨植入钛合金TZS进行喷砂并酸蚀处理对成骨细胞生物学行为的影响。方法采用SD大鼠体外原代培养方法培养成骨细胞，并将细胞分别接种于新型钛合金（ 光滑表面）及表面处理的材料表面，建立体外共同培养模型。采用MTT比色法检测第3天成骨细胞的增殖率，扫描电镜（ SEM）观察细胞形态学变化，培养第5天进行细胞碱性磷酸酶功能活性检测。结果喷砂与酸蚀处理组表面的成骨细胞，在3 d时的细胞增殖率与光滑组比较有统计学差异（ P<0.05）；5 d时的处理组碱性磷酸酶活性检测值与光滑组比较无统计学差异（ P>0.05）；SEM观察成骨细胞在喷砂与酸蚀组表面具有典型形态特征，伸展良好，具有丰富的板状、丝状伪足，优于光滑组。结论喷砂并酸蚀处理的新型钛合金在体外实验表现为不同程度的促进成骨细胞增殖及功能活性表达。     

Toxicity evaluation of root canal sealers in vitro

AIM: To compare the toxicity of methacrylate resin-based root canal sealers with sealers based on epoxy resin and silicone by two-well established cell culture methods. METHODOLOGY: Specimens of AH Plus, EndoREZ, RoekoSeal and Epiphany were prepared for direct contact in the Millipore filter diffusion test and as extracts in the MTT assay. Mouse fibroblasts (L929) were used as toxicity targets. Differences in cytotoxicity between fresh and set specimens and between the extracts of root canal sealers were determined by t-test (P < 0.05). RESULTS: In the filter diffusion test, freshly mixed Epiphany and AH Plus were rated severely toxic and RoekoSeal and EndoREZ nontoxic. When set, Epiphany was moderately toxic, whereas AH Plus, RoekoSeal and EndoREZ were nontoxic. Epiphany was significantly more toxic than RoekoSeal and EndoREZ (P < 0.05). In the MTT assay with set specimens, Epiphany was rated severely toxic; AH Plus and RoekoSeal slightly toxic; and EndoREZ nontoxic. Epiphany was significantly more toxic than the other three materials in this test (P < 0.001). CONCLUSION: The multi-methacrylate resin-based (Epiphany) root canal sealer was significantly more toxic to L-929 cells than the silicone-based Roeko Seal and the single methacrylate-based EndoREZ root canal sealers. AH Plus showed intermediate toxicity.     

新型骨替代材料硅磷酸钙的生物相容性实验研究

目的:评价新型骨替代材料硅磷酸钙的生物相容性,从而为其在骨缺损修复领域中的临床应用提供依据。方法:分别选用含10%、5%硅酸钙的硅磷酸钙,分别进行急性毒性试验、肌肉植入试验。结果:含10%与5%硅酸钙的硅磷酸钙无急性全身毒性,对肌肉组织无刺激,植入肌肉后呈降解趋势。结论:新型骨替代材料硅磷酸钙具有良好的生物相容性。