Viscoelastic properties of acid- and alkaline-treated human dermis: a correlation between total surface charge and elastic modulus

BACKGROUND: One of the major mechanical functions of collagenous tissues is the storage, transmission and dissipation of elastic energy during mechanical deformation. In skin, mechanical energy is stored during loading and then is transmitted and dissipated, which protects skin from mechanical failure. Thus energy storage (elastic properties) and dissipation (viscous properties) are important characteristics of extracellular matrices. METHODS: A uniaxial incremental stress relaxation test method has been used to characterize the time-dependent (viscous) and time-independent (elastic) properties of human dermis. Viscoelasticity was investigated in processed human dermis that was equilibrated at pHs of 3.0, 7.4 and 11.0 in an effort to study the link between electrostatic interactions within the collagen matrix and macroscopic tissue properties. RESULTS: Our results show that the solution pH and the charge on collagen significantly affected the high-strain elastic behavior of dermis; the elastic behavior of skin has previously been shown to be directly correlated with axial stretching of the collagen triple helix in crosslinked collagen fibrils. A positive linear correlation existed between the high-strain elastic modulus and both pH (R(2)=0.96) and the total number of charged residues on collagen (R(2)=0.93). These results provide in vitro/ex vivo evidence that charged groups on the surface of collagen molecules in processed human skin influence the high-strain elastic properties of dermis and are likely to be involved in elastic energy storage. CONCLUSION: It is proposed that the pH and charged residue dependency of the elastic modulus suggests that charged pair interactions and repulsions within and between collagen molecules are involved in elastic energy storage during stretching at high strains. It is hypothesized that elastic energy storage is associated with the stretching of pairs of charged amino acid residues that are found primarily in the flexible regions of collagen molecules.     

脱细胞异体皮肤细胞外基质——医用组织补片在美容外科的应用

目的介绍脱细胞异体皮肤细胞外基质医用组织补片在美容外科中的临床应用经验。方法自2002年3月至2005年10月,应用脱细胞异体皮肤细胞外基质医用组织补片,对患者不同部位进行填充的美容手术,其中头面部填充于皮下浅筋膜层,唇部、颏部、唇裂合并牙槽裂部填充于皮下深筋膜层或肌肉浅层,隆乳术中用于胸大肌减张。结果本组115例患者经1~27个月随访,1例唇裂合并牙槽裂者因补片填充量不足凹陷未完全矫正;1例鼻唇沟填充者其左侧出现血肿、局部出现硬结,于2个月后将左侧假体取出重新填充,但术后6个月仍有不平整,其余患者均获得良好的美容效果。结论脱细胞异体皮肤细胞外基质医用组织补片可以广泛地应用于美容外科的临床填充术。     

Skin ligaments: regional distribution and variation in morphology

Skin ligaments (SL) (L. retinacula cutis) are present extensively in the face, hands, feet, and in breast tissue, but have seldom been reported elsewhere in the body. The traditional histological view of the subcutaneous region is that it comprises a matrix of loose connective tissue devoid of fibrous specializations. The purpose of this study was to determine the structure and distribution of skin ligaments. Eight embalmed cadavers (3 males, 5 females, 69-90 years of age) were used in this study. Tissue was prepared using the E12 plastination technique. Macroscopic and microscopic examination demonstrated the widespread presence in the limbs and most of the rest of the body of fibrous strands linking the base of the dermis and the superficial fibers of the underlying deep fascia. The morphology and distribution of these skin ligaments were similar in the individuals examined. Variations in the structure of the skin ligaments depended on the presence of underlying muscle, neurovascular bundles, intermuscular septa and adipose tissue. We conclude that skin ligaments are complex fibrous structures that are present over most of the body. They form an extensive peripheral network in the subcutaneous fat. These 'ligaments' seem to provide an anchorage of skin to deep fascia that is flexible and yet resistant to mechanical loading from multi-directional forces. The use of the E12 plastination technique coupled with fluorescent confocal microscopy has been of benefit in visualizing and delineating SLs from other soft tissue structures in three planes.     

Ⅰ型胶原膜、胞外基质提取物膜体外构建人工真皮能力比较

比较Ⅰ型胶原与胞外基质(extracellularmatrix,EM)提取物冻干后形成的膜(EM膜)在体外构建人工真皮的能力,以选择一种更有利于人工真皮形成的支架材料。方法分别使用小牛真皮部分提取的Ⅰ型胶原蛋白和胞外基质提取物冻干后制备基质网架,植入皮肤成纤维细胞,形成人工真皮。用扫描电镜观察I型胶原膜和EM膜的表面形态结构;选取不同的时间点,对种植于两种支架材料上的细胞进行计数,并利用间接ELISA方法检测人工真皮复合物中的细胞Ⅰ型胶原分泌情况,通过统计学的分析手段,对细胞的生长情况做出判断;用组织学方法观察人工真皮的形成情况。结果扫描电镜观察结果,两种膜在外观形态上无明显差异;细胞在EM膜上的增殖速度较Ⅰ型胶原膜快;ELISA分析显示,EM-细胞复合物中的成纤维细胞能够分泌更多的Ⅰ型胶原;与Ⅰ型胶原膜相比,EM在培养液中的降解更为缓慢,种植后的成纤维细胞在EM膜上生长旺盛,细胞层次明显多于前者,形成了较为明显的真皮样组织。结论EM膜适于成纤维细胞的生长,体外降解速率慢,是一种较Ⅰ型胶原膜更为理想的真皮支架材料。     

猪脱细胞真皮与自体刃厚皮复合移植的临床应用

目的观察深度烧伤和瘢痕切除创面进行猪脱细胞真皮与自体刃厚皮一步法与两步法复合移植的临床效果。方法2000年4月~2002年12月间用猪脱细胞真皮与自体刃厚皮复合移植修复深度烧伤创面或瘢痕切除创面16例,对一步移植法和两步移植法进行比较。结果一步移植法中,4例植皮完全成活,3例植皮95%以上成活,1例植皮80%成活;两步移植法中,5例植皮完全成活,3例植皮95%以上成活。均无1例补植皮。随访6个月~2年,一步和两步移植法中的复合移植皮肤外观、质地基本接近正常皮肤,瘢痕增生不明显。结论猪脱细胞真皮与自体刃厚皮复合移植是深度烧伤创面、瘢痕切除创面的良好修复材料,一步移植法可行、简便、实用。     

Deletion of mouse Porcn blocks Wnt ligand secretion and reveals an ectodermal etiology of human focal dermal hypoplasia/Goltz syndrome

The Drosophila porcupine gene is required for secretion of wingless and other Wnt proteins, and sporadic mutations in its unique human ortholog, PORCN, cause a pleiotropic X-linked dominant disorder, focal dermal hypoplasia (FDH, also known as Goltz syndrome). We generated a conditional allele of the X-linked mouse Porcn gene and analyzed its requirement in Wnt signaling and embryonic development. We find that Porcn-deficient cells exhibit a cell-autonomous defect in Wnt ligand secretion but remain responsive to exogenous Wnts. Consistent with the female-specific inheritance pattern of FDH, Porcn hemizygous male embryos arrest during early embryogenesis and fail to generate mesoderm, a phenotype previously associated with loss of Wnt activity. Heterozygous Porcn mutant females exhibit a spectrum of limb, skin, and body patterning abnormalities resembling those observed in human patients with FDH. Many of these defects are recapitulated by ectoderm-specific deletion of Porcn, substantiating a long-standing hypothesis regarding the etiology of human FDH and extending previous studies that have focused on downstream elements of Wnt signaling, such as β-catenin. Conditional deletion of Porcn thus provides an experimental model of FDH, as well as a valuable tool to probe Wnt ligand function in vivo.     

Regeneration of nails with artificial dermis

Abstract

We treated fingertips injured through the proximal half of the nail bed using artificial dermis in 22 patients from 2004 to 2009. We classified the injuries to the nail bed into three types according to where the wounds were. Type ? was localised to the nail bed with or without minor injury to the surrounding structure; type II was an avulsion and amputation of the fingertip including the nail bed, the finger pulp, and the distal phalanx at the level of the proximal nail bed; and type III was post-traumatic shortening of the nail, in which the pulp and distal phalanx were intact. Regeneration and elongation of the nail was achieved in every patient by applying artificial dermis. All patients were satisfied with the results.     

Atomic force microscopy for biomechanical and structural analysis of human dermis: A complementary tool for medical diagnosis and therapy monitoring

Skin mechanical properties are usually measured considering the entire skin thickness and very little is known about the mechanical behaviour of individual skin layers. We propose atomic force microscopy (AFM) as a tool to quantify nanoscale changes in the biomechanical properties and ultrastructure of human papillary dermis exposed to different mechanical and physical stimuli. Samples from 3 human skin biopsies were studied: one stretched by obesity, one subjected to a high level of sun exposure and normal skin as control. Slices of the papillary dermis layer were harvested at controlled depths from each skin biopsy and 25 μm² areas of each slice were imaged and D‐periodicity of collagen fibres measured by AFM, together with their stiffness. Standard histological analysis was also carried out to correlate biochemical properties and their distribution with stiffness and topography. We obtained similar stiffness values between the sample affected by obesity and the control sample at any depth level into the dermis, while the sun‐exposed sample presented a significantly lower stiffness. Additionally, all samples presented an increase in the stiffness at higher depths into the papillary dermis layer. Collagen fibres close to the epidermis of sample affected either by obesity and sun exposure—the former even more than the latter—are thicker and present a larger D‐period than those in the control sample. Our results open the possibility to use structural and mechanical analysis based on AFM as a complementary tool for medical diagnosis and therapy monitoring.