Biomechanical researches on tissue engineering bone constructed by deproteinated bone

Objective： To study biomechanical changes of newly formed bones 24 weeks after repairing large defects of long bones of goats using heterogeneous deproteinated bone （DPB） prepared by modified methods as an engineering scaffold. Methods： According to a fully randomized design, 18 goats were evenly divided into three groups： normal bone control group （Group A）, autologous bone group （Group B） and experimental group （Group C）. Each goat in Groups B and C were subjected to the periosteum and bone defect at middle-lower part of the right tibia （20% of the whole tibia in length）, followed by autologous bone or DPB plus autolognus MSCs ＋ rhBMP2 implantation, respectively and semi- ring slot fixation; while goats in Group A did not perform osteotomy. At 24 weeks after surgery, biomechanical tests were carried out on the tibias. Results： At 24 weeks after surgery, the results of anticompression test on tibias in three groups were recorded by a functional recorder presented as linear pressure-deformation curve. The shapes of the curves and their change tendency were similar among three groups. The ultimate pressure values were 10.74 MPa±1.23 MPa, 10. 11 MPa±1.35 MPa and 10.22 MPa±1.32 MPa and fracture compression rates were 26.82%±0.87%, 27.17%±0.75% and 28.22%±1.12% in Groups A, B and C, respectively. Comparisons of anti-compression ultimate pressures and fracture compression rates among three groups demonstrated no significant difference （PAB=0.415, PBC=0.494）. Three-point antibend test on tibias was recorded as load-deformation curves, and the shapes of the curves and their change tendency were similar among three groups. The ultimate pressure values of the anti-bend test were 481.52 N±12.45 N, 478.34 N±14.68 N and 475.62 N±13.41 N and the fracture bend rates were 2.62 mm±0.12 mm, 2.61 mm±0.15 mm and 2.81 mm±0.13 mm in Groups A, B and C, respectively. There was no significant difference between groups （PAB=0.7, PBc=0.448）. The ultimate anti-torsion torque values were 6.55 N.mi-0.25 N.m, 6.34 N＇m^0.18 N＇m and 6.42 N＇m^0.21 N＇m and fracture torsion rates were 29.51°±1.64°, 28.88±1.46° and 28.81°±1.33° in Groups A, B and C, respectively. There was no significant difference between groups （PAB=0.123, PBc=0.346）. Conclusions： The biomechanical characteristics of newly formed bones from heterogeneous DPB for repairing large segmental long bone defect are comparable to those of normal bones and autologous bones. DPB has the potential for clinical usage as bone graft material.

透明质酸钠棉片贴补联合小牛血去蛋白提取物治疗早期创伤性鼓膜穿孔

目的 观察透明质酸钠棉片贴补联合小牛血去蛋白提取物对早期创伤性鼓膜穿孔愈合的促进作用。方法选取南通大学附属医院附属如皋医院2017年1月~2020年1月初诊创伤性鼓膜穿孔病例101例(102耳)。根据患者治疗意愿分为47例治疗组(透明质酸钠棉片贴补联合小牛血去蛋白提取物注射),54例对照组(保守治疗)。比较两组愈合率及愈合时间。结果 治疗组47耳在2周内全部愈合,对照组在2周内仅有6例愈合,16例随访3个月仍未愈合。治疗组在早期创伤性鼓膜穿孔愈合率(P<0.0001)及愈合时间(P<0.0001)均明显好于对照组。鼓膜穿孔面积≤30%和30%~50%的患者,治疗组治愈率(P=0.0216,P=0.0006)和治愈时间(P<0.0001,P<0.0001)均显著优于对照组。鼓膜穿孔面积≥50%的患者(仅8耳,例数较少),治疗组与对照组相比,治愈率(P=0.4286)和治愈时间(P=0.1652)均略优,但两者差距无统计学意义。结论 透明质酸钠棉片贴补联合小牛血去蛋白提取物治疗早期创伤性鼓膜穿孔疗效显著。

Clinical effect of deproteinized calfblood extract eye gel for corneal epithelial defect

Objective: To observe the clinical effect of 20% deproteinized calfblood extract eye gel on the treatment of mechanical injury to corneal epithelium. Methods: 120 cases of patients with corneal epithelial defect caused by mechanical injury were selected and randomly divided into observation group (60 eyes, deproteinized calfblood extract eye gel in use) and control group (0.3% ofloxacin eye drops in use). Both two groups of patients were given eye gel or eye drops 4 times per day. Moreover, the two groups of eye symptoms and corneal epithelial healing were observed on the 3rd day and the 7th day after drug usage respectively. Results: The effective rates of 20% deproteinized calfblood extract eye gel on the treatment of mechanical corneal epithelial defect on the 3rd day and the 7th day were 78.33% and 93.33% respectively. Meanwhile, the effective rates of 0.3% ofloxacin eye drops were 55.00% and 71.67%. The difference in the comparison of the effective rate between two groups was of statistical significance (p < .05). Conclusions: 20% deproteinized calfblood extract eye gel can significantly promote the healing of corneal epithelium in a short time, and it is worthy to be widely applied clinically.

<i>Arthrobacter arilaitensis</i>Re117 as a Source of Solvent-Stable Proteases: Production, Characteristics, Potential Application in the Deproteinization of Shrimp Wastes and Evaluation in Liquid Laundry Commercial Detergents

The present study describes the characterization of crude protease extract from Arthrobacter arilaitensis Re117 and its evaluation in solid and liquid detergent. One caseinolytic protease clear band was observed in zymogram. The crude alkaline protease showed optimum activity at pH 9.0 and 50&degC, and it was highly stable over a wide range of pH from 8.0 to 9.0. Proteolytic enzymes showed extreme stability towards non-ionic surfactants (Tween 80, Tween 20 and Triton X-100) and stimulate activity towards oxidizing agents such as sodium perborate. They also showed high stability and compatibility with various laundry solid detergents from Tunisian market. The protease of A. arilaitensis Re117, was also tested for shrimp waste deproteinization to produce chitin. The protein removal with a ratio E/S of 20 was about 83%. The novelties of the Re117 protease include its high stability to organic solvents and surfactants. These unique properties make it an ideal choice for application in detergent formulations and enzymatic peptide synthesis. In addition, the enzyme may find potential applications in the deproteinization of shrimp wastes to produce chitin.

Properties of deproteinized bone for reparation of big segmental defect in long bone

Objective： To explore suitable scaffold material for big segmental long bone defect by studying the properties of the prepared deproteinized bone. Methods： Cancellated bone were made as 30 mm × 3 mm × 3 mm bone blocks from inferior extremity of pig femur along bone trabecula. The deproteinized bone was prepared with an improved method. Their morphological features, components, cell compatibility, mechanical and immunological properties were investigated respectively. Results： Deproteinized bone maintained natural re-ticular pore system. The main organic material is collagen I and inorganic composition is hydroxyapatite. It has good mechanical properties, cell adhesion rate and histocompatibility. Conlusion： This deproteinized bone can be applicable as scaffold for reparation of big segmental defect in long bone.

Alzheimer's disease treated with combined therapy based on nourishing marrow and reinforcing Qi

OBJECTIVE:To study the efficacy and safety of combined Traditional Chinese Medicine(TCM) therapy based on nourishing marrow to improve intellect and reinforcing Qi to activate bloodon mild to moderate Alzheimer's disease(AD).METHODS:Sixty-six patients with AD,whoseMini-Mental State Examination(MMSE) score were from 10-24,were randomized equally into an intervention group and a control group.The control group was given Aricept(5 mg,once daily).The intervention group was further divided into Yang-Qi deficiency(n = 18) and of Yin-Qi deficiency(n = 15)subgroups.Patients in the Yang-Qi deficiency group were intravenously administered Shenfu injection,60 mL,and deproteinized calf blood injection(DCBl),1.2 g,once daily.The Yin-Qi deficiency group was given Shenmai injection,60 mL,and DCBl,1.2 g,once daily.Each course lasted 21 days.RESULTS:Compared with the control group and with pre-treatment in the same group,MMSE,clinical dementia rating,and activities of daily living scale scores in the intervention group were significantly improved(all P<0.05).These metrics mildly improved in the control group compared with before treatment(P>0.05).No adverse effects were observed in any group during treatment.CONCLUSION:We found that combined TCM therapy is effective and safe for managing mild to moderate AD.

Immunological study on the transplantation of an improved deproteinized heterogeneous bone scaffold material in tissue engineering

Objective： To observe the immune response after the transplantation of a deproteinized heterogeneous bone scaffold and provides the theoretic reference for clinical practice. Methods： The fresh pig bone and deproteinized bone were transplanted respectively to establish BABL/C thigh muscle pouches model of male mice and take the samples for detection at 1, 2, 4, 6 weeks after operation. Lymphocyte stimulation index, subset analysis, serum specific antibody IgG, cytokine detection and topographic histologic reaction after implantation were investigated. Results： After the transplantation of deproteinized bone, lymphocyte stimulation index, CD4^＋ and CD8^＋ T-lymphocyte subsets, serum specific antibody IgG and cytokines in deproteinized bone group were significantly lower than those in fresh pig bone group at each time point （P〈0.05）. The histological examination found that in fresh bone group at each time point, a large quantity of inflammatory cells infiltrated in the surrounding of bone graft, and they were mainly lymphocytes, including macrophages and monocytes. In deproteinized bone group, there were few inflammatory cells infiltration around bone graft one week after operation. The lymphocytes were decreased as time went by. At 6 weeks, fibroblasts and fibrous tissue grew into the graft, and osteoclasts and osteoprogenitor cells appeared on the verge. Conclusions： The established heterogeneous deproteinized bone has low immunogenicity and is a potentially ideal scaffold material for bone tissue engineering.

Improving Energy Metabolism of Deproteinized Extract of Calf Blood Through Regulation of Hmgcs2y Cptla, Angptl4, Cyp8bl, and Ehhadh Genes in Mice

Herein, we described the physicochemical properties of deproteinized extract of calf blood(DECB) and established a hypoxia model treated with or without DECB to detect the sugar, lactic acid, protein, and ATP contents of mice and then identified and analyzed the differentially expressed genes between two groups using mRNA expression chip. According to the results of the airtight hypoxia experiment, mice in the model+DECB group had a signifi? cantly prolonged time of hypoxia tolerance compared with the model group. The biochemical test indicated that DECB could significantly increase the level of sugar, ATP and proteins and reduce the amount of lactic acid in mice. It also revealed that Hmgcs2, Cptla, Angptl4, Cyp8b], and Ehhadh genes were involved in mice energy metabolism, and were closely associated witli metabolic signaling pathway. These results suggest that DECB might be a potential drug to treat metabolic diseases. Among the genes with differential expression under hypoxia, Angptl4, Cyp8bl, and Ehhadh were critical factors for sugar metabolism. Hmgcs2 provided energy directly, and Cptla regulated cellular inflammatory responses, promoting energy metabolism.