铒玻璃点阵激光嫩肤参数优化的动物实验

目的:研究观察铒玻璃点阵激光不同能量密度、微孔密度参数组合方案与嫩肤疗效之间的关系,为临床合理选择治疗参数提供理论参考.方法:清洁级雌性昆明小鼠24只,随机分成4组,每组6只:未照射对照组(A)、高能量密度/低微孔密度组(B)、中能量密度/高微孔密度组(C)、高能量密度/高微孔密度组(D).用1550nm铒玻璃点阵激光照射背部脱毛皮肤,每周1次,共8次.结果:与未照射对照组相比,各照射组真皮厚度、成纤维细胞计数、Ⅰ型及Ⅲ型胶原纤维和弹力纤维容积分数均增加,嫩肤疗效D组＞B组＞C组.结论:1550nm铒玻璃点阵激光嫩肤疗效安全可靠.选择嫩肤治疗参数时,在采用较高能量密度基础上适当提高微孔密度更有利于提高临床疗效.     

点阵CO2激光和Nd∶YAG激光对自然老化小鼠真皮重塑的比较研究

目的:比较点阵CO2激光和Nd:YAG激光(准长脉宽1 064nm波长)对老化昆明小鼠真皮重塑的作用,并探讨二者嫩肤作用机制,为临床选择治疗方法提供理论依据。方法:应用两种波长激光照射自然老化昆明小鼠背部脱毛皮肤,通过HE染色进行成纤维细胞计数及真皮厚度测量,饱和苦味酸-天狼猩红染色观察皮肤Ⅰ型、Ⅲ型胶原纤维的表达及Gomor i醛品红染色观察弹力纤维的变化。结果:两组照射组中真皮厚度、成纤维细胞计数和弹力纤维容积分数的值均高于正常对照组(P均<0.05);点阵CO2激光组均高于Nd:YAG激光(准长脉宽1064nm波长)组(P均<0.05);两组照射组中Ⅰ型及Ⅲ型胶原蛋白容积分数均高于正常对照组(P均<0.05),但照射两组间差别无统计学意义。结论:点阵CO2激光和Nd:YAG激光(准长脉宽1064nm波长)均可引发小鼠真皮重塑,前者效果优于后者。     

内质网应激反应在大鼠Q开关1064nm Nd∶YAG激光非剥脱性嫩肤治疗机制中的作用

目的 探究Q开关1 064 nm Nd∶YAG激光照射对大鼠真皮胶原增生和皮肤中内质网分子伴侣(BiP/GRP78)表达水平的影响,探讨嫩肤治疗中内质网应激反应的作用机制.方法 将25只Wistar大鼠背部皮肤脱毛,平均分成2部分,一部分用Q开关1 064 nm Nd∶YAG激光照射背部脱毛的大鼠皮肤共4次,每次间隔2d(实验组);另一部分不做任何处理(未照射组).取照射后第14和30天的大鼠皮肤组织进行HE染色,测量其真皮厚度;并用碱水解法检测羟脯氨酸含量.取照射后皮肤组织,采用免疫组织化学染色技术检测BiP/GRP78的表达.羟脯氨酸含量和真皮厚度使用t检验,免疫组化使用x2检验,以P ＜0.05为差异有统计学意义.结果 激光照射第14和30天的大鼠皮肤真皮厚度明显增加,增加率分别为29.6％和16.7％,胶原纤维束增粗、密集,皮肤羟脯氨酸含量增加,与未照射组比较差异有统计学意义(P＜0.05).免疫组化显示BiP/GRP78在激光照射后表达率为100％,较未照射组明显上调,差异有统计学意义(x2=28.76,P＜0.01).结论 Q开关1 064 nm Nd∶YAG激光照射大鼠皮肤,引发内质网应激反应,促进蛋白质正确折叠、合成,真皮胶原纤维增粗密集,从而达到非剥脱性嫩肤治疗的目的.     

黄芪延缓小鼠皮肤衰老的实验研究

目的：从形态学角度研究黄芪延缓小鼠皮肤衰老的作用。方法：小鼠随机分成3组,空白组（C）灌胃生理盐水20ml/kg,黄芪低剂量组（L）和黄芪高剂量组（H）分别灌胃黄芪提取液4g/kg、16g/kg,每次20ml/kg。30天后做组织学切片检测。结果：与C组比较,L组和H组皮肤组织中真皮厚度和胶原纤维面积增加（P〈0.01）。H组与L组比较,真皮厚度和胶原纤维面积显著增加（P〈0.05）。结论：高剂量黄芪可使真皮厚度和胶原纤维面积明显增加,具有延缓小鼠皮肤衰老的作用。     

Nd：YAG激光与强脉冲光非剥脱性嫩肤的动物实验

目的探讨Nd：YAG激光长脉宽波长1064nm与强脉冲光（intense pulsed light,IPL）波长560～1200nm两种波长照射SD大鼠皮肤后组织学变化趋势,为临床非剥脱性光嫩肤的科学治疗提供理论依据。方法应用两种波长照射SD大鼠背部脱毛皮肤,通过HE染色和免疫组织化学染色方法检测真皮厚度、Ⅰ型和Ⅲ型胶原蛋白的表达,并测定平均吸光度值进行半定量分析。结果两种波长照射后真皮厚度、Ⅰ型及Ⅲ型胶原蛋白平均吸光度值均高于正常对照组（P〈0．05）,激光组均又高于IPL组（P〈O．05）,Ⅰ型胶原蛋白平均吸光度增加值均高于Ⅲ型胶原蛋白（P〈0．001）。结论两种波长均可使SD大鼠真皮厚度和Ⅰ型、Ⅲ型胶原蛋白增加,激光组优于IPL组。Ⅲ型胶原蛋白在照射早期增加幅度较高,而Ⅰ型胶原蛋白在照射后期增加幅度较高,提示非剥脱性光嫩肤真皮重塑机制可能与Ⅰ型胶原蛋白增加的时间和量有关。     

复合射频除皱嫩肤仪联合胶原蛋白激活生长因子在治疗痤疮及改善肤质中的应用

目的:探讨射频除皱嫩肤仪联合胶原蛋白激活生长因子在治疗痤疮及改善肤质中的确切作用。方法:将2008年3月～2012年12月来笔者科室治疗痤疮并要求改善肤质的72例患者随机分为三组,每组24例。A组:采用复合射频除皱嫩肤仪联合胶原蛋白激活生长因子治疗,并外用消炎、抑制皮脂腺分泌的药物;B组:采用复合射频除皱嫩肤仪治疗,同时使用与A组相同的外用药;C组:仅使用同A组相同的外用药。治疗后随访6～12月,比较各组间的治疗效果。结果:A组治疗痤疮和改善肤质效果明显,B组次之,C组仅有部分治疗痤疮的效果,肤质改善不明显。结论:复合射频除皱嫩肤仪联合胶原蛋白激活生长因子具有较好的治疗痤疮和明显改善肤质的作用,联合使用效用明显增强。     

D980-nm半导体激光对大鼠皮肤溶脂嫩肤作用机制初步研究

目的初步探讨D980-nm半导体激光对SD大鼠皮肤溶脂嫩肤作用机制,分析不同治疗频次的差异。方法12~14月龄清洁级雄性SD大鼠18只,体质量400~450 g。以大鼠脊柱为分界线,左侧皮肤作为激光照射实验侧,右侧皮肤作为自身正常对照侧。实验动物随机分为3组(n=6),实验侧(分别为A、B、C组)采用D980-nm半导体激光仪以脉冲模式,能量密度20 J/cm~2、功率8 W、脉冲宽度20 ms、脉冲频率40 Hz,分别皮下照射1、2、3次作为1个疗程,每次间隔5 min,每个疗程间隔1周,共4个疗程。对照侧(分别为A1、B1、C1组)不作激光处理。第4个疗程结束后1周取材,行HE染色及免疫组织化学染色,观察激光照射前后大鼠皮肤组织结构变化,测量真皮厚度,计数成纤维细胞及TGF-β_1、b FGF阳性细胞。结果与A1、B1、C1组相比,D980-nm半导体激光照射后A、B、C组大鼠皮肤组织结构均改善明显,脂肪细胞减少,局部可见小血管形成,成纤维细胞增多,胶原纤维束增多、变粗且排列紧密、规则,真皮厚度、成纤维细胞以及TGF-β_1、b FGF阳性细胞均显著增加,比较差异有统计学意义(P0.05)。随激光治疗频次的增加,以上指标均逐渐增加,C组优于A、B组,比较差异有统计学意义(P0.05)。结论D980-nm半导体激光治疗对大鼠皮肤有明显的溶脂嫩肤作用,并与激光治疗频次密切相关,其溶脂嫩肤作用与激光的组织效应和细胞效应相关。     

光子嫩肤技术临床疗效的初步观察

目的探讨应用光子嫩肤技术对患者进行颜面皮肤美容的临床疗效初步观察的结果.方法采用光子嫩肤仪某些特定波长的光,选择性地损伤或破坏氧合血红蛋白及黑色素颗粒,刺激纤维细胞分泌新胶原蛋白,从而达到皮肤美容的目的.结果应用光子嫩肤技术对色斑、胶原纤维蜕变、毛细血管扩张的217例患者进行治疗,经4～6次治疗后,痊愈者37例,占17.1%,显效者118例,占54.4%,有效者45例,占20.7%,无效者17例,占7.8%.结论光子嫩肤技术操作简单、损伤小、疗效确切,是一种具有应用前景的新技术.     

光子嫩肤技术临床疗效的初步观察

目的　探讨应用光子嫩肤技术对患者进行颜面皮肤美容的临床疗效初步观察的结果。方法　采用光子嫩肤仪某些特定波长的光 ,选择性地损伤或破坏氧合血红蛋白及黑色素颗粒 ,刺激纤维细胞分泌新胶原蛋白 ,从而达到皮肤美容的目的。结果　应用光子嫩肤技术对色斑、胶原纤维蜕变、毛细血管扩张的 2 1 7例患者进行治疗 ,经 4～ 6次治疗后 ,痊愈者 37例 ,占 1 7.1 % ,显效者 1 1 8例 ,占 54 .4% ,有效者 45例 ,占 2 0 .7% ,无效者 1 7例 ,占 7.8%。结论　光子嫩肤技术操作简单、损伤小、疗效确切 ,是一种具有应用前景的新技术     

双极射频治疗山羊膝关节软骨损伤的实验研究

目的 探讨双极射频在治疗关节软骨损伤中的价值.方法 采用前交叉韧带切断和内侧半月板切除的方法制造16只山羊股骨软骨继发性损伤的动物模型.3个月后采用双极射频对右膝的损伤软骨进行治疗,左膝为对照侧.观察双极射频对损伤软骨的即刻效应和后期效应.结果 造模手术造成以股骨内侧髁软骨Outerbridge Ⅱ级和Ⅲ级为主的损伤.即刻观察显示射频处理后的软骨表面光整,小的裂隙被融合.浅层的少量软骨细胞死亡,深层细胞受到的影响比较小.3个月后发现治疗侧股骨髁软骨表面仍光滑,改良Mankin's软骨评分结果显示治疗侧(12±4)分,对照侧(14±5)分(P＜0.01),治疗侧软骨损伤比对照侧轻.流式细胞学检测发现治疗侧软骨细胞死亡率(24±10)%,对照侧(12±1)%,两组比较差异无统计学意义(P＞0.05).结论 一个能级的双极射频不会对软骨细胞造成明显损害,软骨成形术是治疗软骨损伤的有效方法之一.     

生物支架材料胶原膜交联前后的特性分析

目的 比较胶原膜交联前后的生物特性,为胶原膜的应用奠定基础. 方法 取市售新鲜牛尾,分离肌腱,制备胶原蛋白,以紫外分光光度计测定胶原蛋白含量;采用SDS-聚丙烯酰胺凝胶电泳(SDS-polyaerylamide gelelectrophoresis,SDS-PAGE)波长扫描,氨基酸含量检测分析胶原蛋白特性.将制备的胶原蛋白,真空冷冻干燥,制备胶原膜,采用戊二醛交联.大体观察及扫描电镜观察交联前后胶原膜形态,并测定胶原酶溶解时间、pH值、吸水性能及抗撕强度.取自愿捐赠的骨髓制备BMSCs,按2×103/100 μL浓度接种于交联前后胶原膜上培养7 d,测量细胞吸光度(A)值及扫描电镜观察. 结果 制备的胶原蛋白含量约为2 mg/mL,最大吸收波长约为230 nm.胶原蛋白氨基酸分析显示,甘氨酸含量约为21.47%,脯氨酸12.04%,羟脯氨酸10.18%,未检出色氨酸.交联前胶原膜为白色海绵状,孔径较大且不均匀,pH值为4.5～5.0;扫描电镜观察为孔网状结构,孔径大;吸水力为其重量的61倍,抗撕强度为210 g/cm3,胶原酶溶解时间为30 min.交联后胶原膜变薄,无色,半透明,致密,柔韧性好;镜下可见致密排列的胶原纤维成网状;pH值为6.5～7.0,吸水力降低,抗撕强度约为3 400 g/cm3,胶原酶溶解时间为90 min.细胞增殖实验表明,细胞在交联前后的胶原膜上均能良好生长,A值差异无统计学意义(P＞0.05).扫描电镜见BMSCs在交联后的胶原膜上生长良好. 结论 从牛肌腱中成功提取胶原蛋白,并制备胶原蛋白膜.经化学交联剂交联后,胶原膜仍具有良好的生物学特性,且理化性质优于交联前,可作为生物支架材料,用于皮肤创伤修复等研究领域.     

The wound healing potential of collagen peptides derived from the jellyfish Rhopilema esculentum

Purpose: Wound represents a major health challenge as they consume a large amount of healthcare resources to improve patient''s quality of life. Many scientific studies have been conducted in search of ideal biomaterials with wound-healing activity for clinical use and collagen has been proven to be a suitable candidate biomaterial. This study intended to investigate the wound healing activity of collagen peptides derived from jellyfish following oral administration. Methods: In this study, collagen was extracted from the jellyfish–Rhopilema esculentum using 1% pepsin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fourier transform infrared (FTIR) were used to identify and determine the molecular weight of the jellyfish collagen. Collagenase II, papain and alkaline proteinase were used to breakdown jellyfish collagen into collagen peptides. Wound scratch assay (in vitro) was done to determine migration potential of human umbilical vein endothelial cells (HUVEC) covering the artificial wound created on the cell monolayer following treatment with collagen peptides. In vivo studies were conducted to determine the effects of collagen peptides on wound healing by examining wound contraction, re-epithelialization, tissue regeneration and collagen deposition on the wounded skin of mice. Confidence level (p < 0.05) was considered significant using GraphPad Prism software. Results: The yield of collagen was 4.31%. The SDS-PAGE and FTIR showed that extracted collagen from jellyfish was type I. Enzymatic hydrolysis of this collagen using collagenase II produced collagen peptides (CP1) and hydrolysis with alkaline proteinase/papain resulted into collagen peptides (CP2). Tricine SDSPAGE revealed that collagen peptides consisted of protein fragments with molecular weight <25 kDa. Wound scratch assay showed that there were significant effects on the scratch closure on cells treated with collagen peptides at a concentration of 6.25 mg/mL for 48 h as compared to the vehicle treated cells. Overall treatment with collagen peptide on mice with full thickness excised wounds had a positive result in wound contraction as compared with the control. Histological assessment of peptides treated mice models showed remarkable sign of re-epithelialization, tissue regeneration and increased collagen deposition. Immunohistochemistry of the skin sections showed a significant increase in b-fibroblast growth factor (b-FGF) and the transforming growth factor-b1 (TGF-b1) expression on collagen peptides treated group. Conclusion: Collagen peptides derived from the jellyfisheRhopilema esculentum can accelerate the wound healing process thus could be a therapeutic potential product that may be beneficial in wound clinics in the future.     

A case of bilateral inguinal hernia recurrence in infancy: Investigations on collagen metabolism

Background. Recurrent inguinal hernias in early infancy are rare. We report on a case of a 3-month-old male infant suffering bilateral inguinal hernia recurrence (RINGH). Due to previous observations of an altered collagen metabolism in hernia patients, a severe connective-tissue pathology in the infant was hypothesised. Methods. Hernial sac tissue of the infant was analysed and compared to specimens from five children operated upon one-sided primary inguinal hernias (controls). In paraffin-embedded sections, we determined the distribution of collagen types I and III by crosspolarisation microscopy and the expression of matrix metalloproteinase 2 (MMP-2) by immunohistochemistry. In fibroblast cultures, expression of collagen types I and III and of MMP-2 was investigated by RT-PCR (real-time polymerase chain reaction) and zymography. Electron microscopical investigations were performed exemplarily in two fibroblast cultures to compare cell morphology. Results. No differences in collagen I/III ratios between RINGH and controls were found either on protein or on mRNA level. Immunohistochemical and RT-PCR analysis of MMP-2 showed a lowered expression in the RINGH patient, as compared to controls, whereas the gelatinolytic activity of MMP-2 did not differ between the groups. Electron microscopical investigations showed similar cell arrangement and morphology. Conclusions. To conclude, a marked biochemical correlate to a severe connective-tissue pathology in the infant suffering inguinal hernia recurrence could not be found. With regard to the slight differences in the expression of MMP-2, a possible role in the genesis of inguinal hernia recurrence cannot be ruled out.     

椎体终板损伤对兔椎问盘髓核Ⅰ、Ⅱ型胶原的影响

目的 观察椎体终板损伤对兔椎间盘髓核Ⅰ、Ⅱ型胶原的影响并探讨其机制.方法 4个月龄清洁级日本大白兔12只,完整取出40个包含终板的完整椎间盘(L2-L5),随机分为实验组和对照组,每组20个.实验组参考Daniel Haschtmann的方法建立终板损伤模型.椎间盘整体培养2周后,免疫组织化学方法观察椎间盘髓核中Ⅰ、Ⅱ型胶原的表达情况,并用HMIAS-2000图像分析系统进行半定量分析.结果 免疫组织化学染色显示:实验组椎间盘髓核Ⅰ型胶原的表达比对照组高;而Ⅱ型胶原的表达则比对照组低,差异均有统计学意义(P<0.05).结论 外伤致椎体终板损伤后会导致椎间盘髓核Ⅰ、Ⅱ型胶原的变化,继而加速椎间盘退行性变.     

Piezoelectricity in collagen films

Summary Tissue collagen exhibits several levels of structural organization, and this complicates efforts to determine the origin of its piezoelectricity. We made collagen films—by evaporation and electrodeposition from solution—and examined the relation between collagen's piezoelectricity and its electron microscopic appearance. We found that the electrodeposited films were more organized and exhibited higher piezoelectric coefficients than the evaporated films. Despite this, the evaporated films were piezoelectric, thereby suggesting that the effect originates either at the level of the tropocollagen molecule or, at most, with aggregated structures no larger than 50 ? in diameter.     

诱导性人工骨膜的研制及实验研究

目的　本研究研制了一种诱导性人工骨膜。方法　用猪真皮提取纯化胶原,盐析法制备胶原膜,并用此吸附ＢＭＰ,制备成胶原－ＢＭＰ人工骨膜。用大鼠桡骨３ｍｍ长骨缺损作为骨折模型。设空白、单纯胶原膜、胶原－ＢＭＰ膜包绕骨折段以及新鲜同种异体植骨共４组。术后０．５、１、２和３个月分批处死各组大鼠数只,处死前２天每只大鼠腹腔内注射钙黄绿素８０ｍｇ／ｋｇ。处死后分别行Ｘ线和软Ｘ线摄片,血清ＢＧＰ测定以及荧光显微镜观察。结果　胶原－ＢＭＰ人工骨膜组的上述多项检测指标均表明其成骨效能明显优于其他各组。结论　作者认为该组成骨效果优良的原因是由于胶原膜吸附了ＢＭＰ,同时具有骨诱导和骨引导的两种成骨作用,而仅胶原膜包绕骨折段的单纯骨引导效果并不理想。     

Analysis of collagens solubilized from cartilage of normal and spontaneously osteoarthritic rhesus monkeys

Osteoarthritis (OA) is a disorder which results in the destruction of the articular cartilage and the remodeling of the subchondral bone in synovial joints. We have analyzed the cartilage collagen from normal and osteoarthritic free-ranging rhesus monkeys from the Cayo Santiago colony. The cartilage samples were assigned a severity score based on histological staging system and were divided into four groups (normals, mild OA, moderate OA and severe OA). After a 4.0 M guanidinium chloride (GuCl) extraction, the remainder of the cartilage was digested with pepsin and the collagen was salt precipitated at 2.5 M and 4.3 M NaCl. The GuCl solubility of the osteoarthritic cartilage increased compared to normals. Collagen extractability by GuCl also increased with the severity of disease. Pepsin digestion followed by salt precipitation shows that collagen from rhesus osteoarthritis cartilage is more easily extracted than from normal cartilage. With an anti-type I collagen antibody we have detected the presence of type I collagen in the severe OA cartilage samples but not in the milder OA groups or in normal cartilage. Total collagen content decreases with severity of OA, which is not due to changes in propyl hydroxylation because examination of collagen hydroxylation, based on hydroxyproline analysis, shows no difference between OA and normal cartilage.     

BSAVA Manual of Canine and Feline Anaesthesia and Analgesia, 2nd Ed.

The late appearance of incisional hernias several years after laparotomy and the high recurrence rates after operation strongly imply the presence of a disorder of the connective tissue, although a specific defect in patients with incisional hernias has not yet been identified. In the present study we used both immunohistochemistry and Western blot analysis to evaluate the ratio of collagen I and III and the expression of the metalloproteinases (MMP) 1 and 13 in the fascia of patients with incisional or recurrent incisional hernias. Samples of healthy skin or stable skin scar in patients without hernias served as controls. Altogether, our data indicated a significantly decreased ratio of collagen I/III in the fascia of patients with incisional hernias and recurrent incisional hernias. Furthermore, in these patients the expression of MMP-1 was decreased compared to the controls, whereas MMP-13 could not be detected in any fascia sample, with or without hernias present. For the first time, our results give evidence of the existence of a possible collagen disorder in these patients. The decreased ratio of collagen I/III is explainable due to a relative increase of collagen type III, which is known to be characterized by thin fibril diameters and lowered mechanical strength. The altered collagen ratio might be the result of the decreased activity of MMP-1, whereas the absent MMP-13 expression did not seem to modify the scar formation. Thus, our data indicate the presence of collagen metabolic disorders in patients with incisional hernias and recurrent incisional hernias. Furthermore, these results might explain the poor results of a mesh-free hernia repair, which again builds up scar tissue of inadequate collagen composition and strength.