Employing optical microscopy technique to determine the tissue action of Hamamelis virginiana L. (Hamamelidaceae), Maytenus ilicifolia Mart. ex Reissek (Celastraceae) and Casearia sylvestris Sw. (Salicaceae) tinctures on skin-fixed histological sections of Wistar rats

Objective: In this study, we aimed to determine the action of the tinctures of Hamamelis virginiana, Maytenus ilicifolia, and Casearia sylvestris on tissues. For this, we investigated the histological sections of fixed skin tissue of Wistar rats for the changes in collagen and elastic fibers, epithelial cells, conjunctive cells, epidermal attachments, pigments, and granules using the optical microscopy technique. Since in the literature and published articles, the use of in vivo models, such as Wistar rats, is predominant to evaluate the healing action of herbal medicines. Methods: The tinctures of Hamamelis virginiana, Maytenus ilicifolia, and Casearia sylvestris, and ethanol 70% (blank) were used. The tinctures were obtained at 10% (w/v) through percolation using 70% ethanol (v/v) as the extraction liquid. This study was conducted in duplicate for each tincture and different incubation times (4, 24, and 48 h) at 37 °C in an oven. The slides used in this study (Wistar rat skin) were previously processed at the histology laboratory since the waste material was donated to conduct this experiment. The Research Ethics Committee approved the use of animals of the Life Sciences Center of the Pontifical Catholic University of Campinas (PUC-CAMPINAS) under the protocol approval number 367/08. Before initiating the staining process, the slides were removed from the oven, and the tincture deposited on the slides was removed. Weigert staining was performed subsequently. For semi-quantitative analysis, the histological sections were carefully observed, and the number of collagen and elastic fibers was evaluated based on the following scale: (+) normal fiber presence, (−) slight decrease, (− −) moderate decrease, and (− − −) intense decrease. The images were digitally captured to obtain the results using a photomicroscope. Results: The degradation of collagen fibers was best evidenced upon using Hamamelis virginiana tincture, which is concordant with the existing reports in the literature on its healing action via the precipitation of dermal proteins. Maytenus ilicifolia and Casearia sylvestris tinctures exhibited low proteolytic capacity as they only caused degradation of elastic fibers that are more delicate and very different from collagen in their constitution. Therefore, the application of the latter two as healing agents (which acts through precipitating proteins) is limited. Conclusions: This experimental histological model using the optical microscopy technique exhibits the advantage of agility and high efficiency as a simple and powerful approach.

Oxidative stress is involved in LLLT mechanism of action on skin healing in rats

The skin injury healing process involves the main phases of homoeostasis, inflammation, proliferation, and remodeling. The present study aimed to analyze the effects of low-level laser therapy (LLLT) on hematological dynamics, oxidative stress markers, and its relation with tissue healing following skin injury. Wistar rats were divided into control, sham, skin injury, and skin injury LLLT. The biochemical and morphological analyses were performed in the inflammatory (1 and 3 days) and regenerative phases (7, 14, and 21 days) following injury. The skin injury was performed in the dorsal region, between the intrascapular lines, using a surgical punch. LLLT (Al-Ga-In-P, λ=660 nm, energy density of 20 J/cm2, 30 mW power, and a time of 40 s) was applied at the area immediately after injury and on every following day according to the experimental subgroups. LLLT maintained hematocrit and hemoglobin levels until the 3rd day of treatment. Surprisingly, LLLT increased total leukocytes levels compared to control until the 3rd day. The effects of LLLT on mitochondrial activity were demonstrated by the significant increase in MTT levels in both inflammatory and regenerative phases (from the 1st to the 7th day), but only when associated with skin injury. The results indicated that LLLT modulated the inflammatory response intensity and accelerated skin tissue healing by a mechanism that involved oxidative damage reduction mostly at early stages of skin healing (inflammatory phase).

Discovery and molecular analysis of conserved circRNAs from cashmere goat reveal their integrated regulatory network and potential roles in secondary hair follicle

Background: Circular RNAs, a novel class in the eukaryotic transcriptome, are characterized by the 3' and 5' ends that are covalently joined in a covalently closed loop without free ends. Circular RNAs are considerably stable molecules and act as microRNA sponges with regulatory potential to the protein-coding genes. Results: Eight circular RNAs were found to be significantly upregulated at anagen skin tissue of cashmere goat compared with their counterparts at telogen. Rich and complex regulatory patterns were revealed among the eight upregulated circular RNAs at anagen and related miRNAs with their potential regulatory genes. The potential regulatory genes of eight upregulated circular RNAs at anagen were involved in several pathways related to the main physiological process of hair follicle, such as histone acetylation and axon. For chi_circ_1926, chi_circ_3541, chi_circ_0483, chi_circ_3196, and chi_circ_2092, overall, the relative expression in secondary hair follicle exhibited highly similar trends with their corresponding host genes during the different stages of the hair follicle cycle. However, the expression trends of chi_circ_0100, chi_circ_2829, and chi_circ_1967 were found to diverge from their corresponding host genes during the different stages of the hair follicle cycle. Conclusions: A total of eighteen circular RNAs were identified and characterized from skin tissue of cashmere goat. The eight upregulated circular RNAs at anagen might have significant roles in the secondary hair follicle of cashmere goat. Our results would provide a novel regulatory layer to elucidate the molecular mechanisms underlying the development of secondary hair follicle and the growth of cashmere fiber in cashmere goat.

Screening of Sera from Patients with Pancreatitis by an Apoptosis Assay of Skin-derived Cells / 대한소화기학회지

BACKGROUND/AIMS: An excessive inflammatory response is typical in acute pancreatitis and a significant cause of early mortality in severe acute pancreatitis. This is believed to be caused by inflammatory molecules or upregulated cytokine levels in the serum of patients. The aim of this study was to identify the serum-mediated apoptosis-inducing effects in acute pancreatitis patients.METHODS: A skin tissue-derived cell line, BJ, was treated for 24 hours with the sera of 22 healthy volunteers (control) and 71 acute pancreatitis patients (22 with gallstone pancreatitis, 16 with alcoholic pancreatitis, and 11 with pancreatitis with other causes) collected at the time of hospital admission (active) and discharge (resolved). Apoptosis was analyzed by flow cytometry.RESULTS: The average percentage of living cells, early apoptotic cells, and late apoptotic cells ranged from 78.8% to 85.0%, 5.5% to 7.3%, and 7.7% to 13.1%, respectively. The number of live cells increased significantly using the serum from the resolved state of gallstone-induced pancreatitis. In addition, the number of early apoptotic cells increased significantly using the serum from the resolved state of pancreatitis with other causes. The number of late apoptotic cells decreased significantly with the serum from the resolved state compared to the active state of gallstone- and alcohol-induced pancreatitis.CONCLUSIONS: Serum samples from patients with pancreatitis induced a change in the apoptosis profiles of skin-derived cells. These results indicate changes in the serum components in patients with acute pancreatitis.

Screening of Sera from Patients with Pancreatitis by an Apoptosis Assay of Skin-derived Cells / 대한소화기학회지

BACKGROUND/AIMS: An excessive inflammatory response is typical in acute pancreatitis and a significant cause of early mortality in severe acute pancreatitis. This is believed to be caused by inflammatory molecules or upregulated cytokine levels in the serum of patients. The aim of this study was to identify the serum-mediated apoptosis-inducing effects in acute pancreatitis patients. METHODS: A skin tissue-derived cell line, BJ, was treated for 24 hours with the sera of 22 healthy volunteers (control) and 71 acute pancreatitis patients (22 with gallstone pancreatitis, 16 with alcoholic pancreatitis, and 11 with pancreatitis with other causes) collected at the time of hospital admission (active) and discharge (resolved). Apoptosis was analyzed by flow cytometry. RESULTS: The average percentage of living cells, early apoptotic cells, and late apoptotic cells ranged from 78.8% to 85.0%, 5.5% to 7.3%, and 7.7% to 13.1%, respectively. The number of live cells increased significantly using the serum from the resolved state of gallstone-induced pancreatitis. In addition, the number of early apoptotic cells increased significantly using the serum from the resolved state of pancreatitis with other causes. The number of late apoptotic cells decreased significantly with the serum from the resolved state compared to the active state of gallstone- and alcohol-induced pancreatitis. CONCLUSIONS: Serum samples from patients with pancreatitis induced a change in the apoptosis profiles of skin-derived cells. These results indicate changes in the serum components in patients with acute pancreatitis.

The Incorrect Translation, Misinterpretation and Misunderstanding of "Superficial Fascia" in Acupuncture / 针刺研究

The "superficial fascia" is often interpreted as "tendon-membrane like structure" in acupuncture field and it is speculated that acupuncture stimulation-induced structural alteration of the superficial fascia may contribute to the alleviation of many medical conditions. The fact is that the superficial fascia mainly consists of subcutaneous adipose tissue and fibrotic network (which function in storing, supporting and buffering external pressure and are not responsible for producing "Deqi" or "needling sensations"), and has no typical tendon or membrane structure in hypodermis. In the present paper, the author tries to clarify its incorrect translation, misinterpretation, and misunderstanding in acupuncture and propose that the target of acupuncture stimulation is probably not only the subcutaneous soft tissue per se, but also the contents in subcutaneous soft tissue network as the nerves, blood vessels, lymph-vessels, cutaneous accessory structures, mast cells, multiple movable cells, related receptors, etc.

Application of high-frequency ultrasound in dermabrasion of patients with deep partial-thickness burns / 中华烧伤杂志

Objective@#To investigate the application of high-frequency ultrasound in dermabrasion of patients with deep partial-thickness burns.@*Methods@#Twenty-six patients with deep partial-thickness burns conforming to the study criteria were hospitalized in our unit from March 2015 to March 2016. Patients were all performed with dermabrasion. The structure of skin tissue and blood flow signals of uninjured side and wounds before dermabrasion, and those of wounds immediately post dermabrasion and on post dermabrasion day (PDD) 1, 3, 5, 7, 10, 14, and 21 were detected with high-frequency ultrasound, and the percentage of blood flow signals was calculated. According to the results of comparison between percentage of blood flow signals of wounds and that of normal skin before dermabrasion, patients were divided into no significant decrease group (NSD, n=19) and significant decrease group (SD, n=7). Wound healing time of patients in two groups was recorded. Data were processed with analysis of variance of repeated measurement, LSD test, t test and Chi-square test. The correlation between the percentage of blood flow signals of wounds before dermabrasion and wound healing time of 26 patients were analyzed by Spearman correlation analysis.@*Results@#(1) Epidermis of normal skin of patients in two groups before dermabrasion showed continuous smooth linear hyperecho, which was stronger than that of dermis, and boundary of dermis and subcutaneous tissue showed stronger discontinuous linear echo than that of dermis, which gradually transited to subcutaneous tissue. In group NSD, epidermis of wound of patients before dermabrasion showed intermittent rough linear echo, which was weaker than that of normal skin epidermis, and there was no obvious abnormity of boundary between dermis and subcutaneous tissue. Immediately post dermabrasion and on PDD 1, no linear hyperecho of epidermis was observed, showing complete attrition of epidermis, and the echo of dermis and subcutaneous tissue had no obvious change as compared with that before dermabrasion, with flat surface of dermis and partly abraded superficial-dermis but relatively well preserved dermal tissue in whole. The epidermis showed discontinuous linear hyperecho, and epidermis was discontinuously regenerated on PDD 3 and 5. Partial continuous linear hyperecho was detected in the epidermis, showing partial continuous regeneration of epidermis on PDD 7 and 10. The regenerated epidermis was thicker than normal skin epidermis and showed rough linear hyperecho with non-uniform thickness on PDD 14. The regenerated epidermis was thicker than normal skin epidermis and showed rather smooth linear hyperecho with uniform thickness on PDD 21. In group SD, the structure of epidermis and dermis of wound of patients before dermabrasion, immediately post dermabrasion, and on PDD 1 was similar to that in group NSD, but the echo of boundary of dermis and subcutaneous tissue was weakened in different degrees. There was no linear hyperecho of epidermis, showing no epidermis was regenerated on PDD 3 and 5. Intermittent regeneration of epidermis appeared on PDD 7 and 10 with intermittent linear hyperecho. Partial continuous linear hyperecho was detected in the epidermis, showing partial continuous regeneration of epidermis on PDD 14 and 21. (2) The percentages of blood flow signals of wounds of patients in group NSD before dermabrasion, immediately post dermabrasion, and on PDD 1 were (3.1±1.3)%, (6.5±2.0)%, and (5.3±1.9)% respectively, higher than those in group SD [(0.9±1.1)%, (3.5±1.3)%, and (3.6±0.9)% respectively, P<0.05 or P<0.01]. The percentages of blood flow signals of wounds of patients in two groups were similar at the other time points (with P values above 0.05). Compared with the percentage of normal skin in the same group [(3.2±0.7)%], the percentages of blood flow signals of wounds of patients in group NSD were significantly increased immediately post dermabrasion and on PDD 1 (with P values below 0.01) but had no significant change at the other time points (with P values above 0.05). The percentage of blood flow signals of wounds of patients before dermabrasion in group SD was significantly lower than that of normal skin in the same group [(2.8±0.6)%, P<0.01]. The percentage of blood flow signals of wounds of patients in group SD was close to that of normal skin in the same group at each time point post dermabrasion (with P values above 0.05). (3) The wound healing time of patients in group NSD was (16.2±2.5) d, lower than that in group SD [(30.9±2.9) d, t=12.67, P<0.01]. There was obvious negative correlation between the percentage of blood flow signals of wounds before dermabrasion and wound healing time of 26 patients (r=-0.77, P<0.01).@*Conclusions@#High-frequency ultrasound is a good way to observe the imaging features of wounds in patients with deep partial-thickness burns before and after dermabrasion, and it can provide objective imaging evidence for the performance of dermabrasion in patients with deep partial-thickness burns.

Progression of related diseases of desmoglein abnormal and pharmacological intervention / 中国药理学通报

Desmoglein,which is a kind of Ca2+ dependent desmosomal cadherins protein,is a major part of the desmosomes.The desmoglein that distributes in the epithelium,myocardium and other tissues plays a very important role in the cell junction.In recent years,the detection of the abnormal expression and function of the desmoglein was proved in many diseases,such as skin,mucous membrane and tumor related diseases.Drugs may have an important effect in the treatment of diseases by interfering with the expression and function of desmoglein.In this paper,the distribution of several subtypes of the family of desmosomes and their functions in the related diseases are reviewed,which may also provide some new clues for the new drug research on the target of desmogleins.

Tissue expansion in treatment of facial congenital giant nevus / 中华医学美学美容杂志

Objective To investigate the effect of tissue expansion in facial congenital giant nevus.Methods From October 2014 to October 2016,7 cases of facial congenital giant nevus patients were treated with expanded skin flaps,including single expansion and secondary expansion.Partial excision and skin tissue expansion were used to drastically remove giant pigmented nevi on body or extremities of infants.There were some points for attention during the surgery:the size and location of the partial excision and expander should be designed reasonably;no tumor-manipulation and non-tension principle should be taken great notice,and expanders should be linked up with one another when several expanders were used at the same time,so as to reduce the possibility of forming the envelop,which was favorable for the second surgery of transfer of a skin flap.Results All cases of giant pigmented nevi excision were on the face,which were completely removed from the patients.The areas of the nevi were from 7 cm × 5 cm to 15 cm × 13 cm.All patients were followed up for 3-12 months (mean 6 months).All flaps survived well.Expander exposure was happened in 2 cases with no bad results.One case of scar received late repair due to surgical scars and all got satisfactory results.Conclusions Tissue expansion is an ideal method in the treatment of facial congenital giant nevus.

Nursing of patients undergoing skin tissue biopsy on head and face / 现代临床护理

Objective To summarize the experience in nursing patients undergoing skin biopsy on head and face . Method One hundred and twenty patients with head facial dermatosis undergoing skin tissue biopsy were nursed from August 2012 to October 2013. Result All operation for biopsy were completed smoothly, with the cure rate at the first phase 100.0%, healing time 5~7 days, averaged (7.0 ± 3.0)d and no complications. Conclusion For those patients having undergone facial skin tissue biopsy, the nursing measures like psychological nursing, health education before operation, preoperative relief of mental disorder, postoperative wound nursing and health education, prevention of complications are critical for their recovery.

Effect of JNK expression in dorsal root ganglia on histological change of foot skin in diabetic rats / 中国病理生理杂志

[ ABSTRACT] AIM:To observe the change of skin histology in diabetic rats and to investigate the possible me-chanism of c-Jun N-terminal kinase (JNK) protein in the dorsal root ganglion (DRG) during the process.METHODS:Diabetic animal model was established in the male SD rats by intraperitoneal injection of streptozotocin.Plantar skin speci-mens of the rats were collected from control group, DM 2-week group (DM2), DM 4-week group (DM4), and DM 8-week group ( DM8) .Immunohistochemical staining and HE staining were used to observe the change of PGP 9.5 immunoreactive nerve terminals and the structures of the skin tissues.The protein expression of PGP 9.5 in the plantar skin tissues, and JNK and p-JNK protein in the DRG within lumbar 5, 6 (L5, 6), and sacral 1 (S1) spinal cord segments were detected by Western blotting.RESULTS:PGP 9.5 immunoreactive nerve terminals of the plantar skin of the rats mainly distributed in the basal layer of the epidermis and papillary dermis.Compared with control group, PGP 9.5 positive nerve terminals in DM4 group showed reduced density and sparse distribution.PGP 9.5 positive nerve terminals in DM8 group showed signifi-cantly reduced distribution, thinner nerve diameter, shorter length and distorted shape.Histological changes of the thinner epidermal tissue, reduced epidermal cell layers, uneven cell distribution and arrangement in DM4 group, and significantly reduced epidermal cell layers, swollen and blurred cells, increasing cell gap, lack of stratified epidermis arrangement for part of epidermis, atropal and degenerated dermal collagen fiber, significantly decreased subcutaneous fat in DM8 group were observed.The results of Western blotting showed that the protein expression of PGP 9.5 in the plantar skin tissue of DM rats was progressively decreased along with the disease, while the protein level of p-JNK in L5, 6-DRG or S1-DRG showed a gradual increasing trend.PGP 9.5 immunoreactive positive nerve terminal density of plantar skin in DM rats had a negative correlation with the protein level of p-JNK in L5, 6-DRG and S1-DRG (P<0.01), but showed a significant positive correlation with the plantar skin thickness (P<0.01).CONCLUSION:The protein level of p-JNK within L5, 6-DRG or S1-DRG in DM rats shows a progressive enhancement.At the same time, there is a significant change in the skin tissue density and structure.The changes of skin tissue and nerve morphology in DM rat may be related to the activation of JNK/SAPK pathway in L5, 6-DRG or S1-DRG cells.Blocking or inhibiting JNK/SAPK pathway may delay the diabetic pe-ripheral neuropathy and reduce the risk of skin lesions.

Curative effect observation on anterolateral calf flap with extra-territorial blood supply in repairing large area of foot skin tissue injury / 局解手术学杂志

Objective To study the effect of anterolateral calf flap with extra-territorial blood supply in repairing large area of foot skin tissue injury. Methods 40 cases of arge area of foot skin tissue injury were divided into the observation group which were given anterolateral crural flap transplantation and the control group which were given anterolateral thigh flap transplantation. The wound healing condition and walking ability were observed. Results The level of wound healing, Holden walking ability and FAC walking ability were better than control group (P<0. 05). In the observation group, wound healing time was (15. 2 ± 6. 2) d and donor skin crusting time was (9. 4 ± 1. 5) d, which were less than the control group(P<0. 05). Conclusion Anterolateral calf flap with extra-territorial blood supply has positive clinical value as it can improve wound healing and walking function.

Gene expression of the skin and sweat glands in different development stages of embryo / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.24.008

Notoginseng glycosides effects on hyperplastic scar / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.24.010

Photochemical tissue bonding in tissue repair / 上海交通大学学报（医学版）

Suture was a traditional method for tissue repair. However, sutures used in the closure of surgical wounds can induce inflammation and lead to scarring that appears as crosshatch marks. Threads used for sutures may cause immune response and surgical infection. In this review, we introduce a novel sutureless technique for wound repair, called photochemical tissue bonding (PTB). Absorption of visible light by a photosensitizing dye initiates photochemical reactions leading to covalent protein-protein crosslinks that bridge the wound surfaces, thus producing an immediate seal between the wound surfaces. This technique has been applied in various tissues including skin, cornea, nerve, blood vessels, et al. PTB reduces inflammation resulting in less scarring in skin. PTB dramatically improves the incidences of vascular anastomotic leakage and stricture. In addition, PTB enhances the recovery of nerve function after nerve repair.

Quantification of thermal damage in skin tissue / 药物分析学报

Skin thermal damage or skin burns are the most commonly encountered type of trauma in civilian and military communities. Besides, advances in laser, microwave and similar technologies have led to recent developments of thermal treatments for disease and damage involving skin tissue, where the objective is to induce thermal damage precisely within targeted tissue structures but without affecting the surrounding, healthy tissue. Further, extended pain sensation induced by thermal damage has also brought great problem for burn patients. Thus, it is of great importance to quantify the thermal damage in skin tissue. In this paper, the available models and experimental methods for quantification of thermal damage in skin tissue are discussed.

Imbalance of MMP-9/TIMP-1 during the cutaneous "underlying disorder" in diabetic rats / 中华内分泌代谢杂志

Objective To investigate the change of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the skin of diabetic rats, and to explore the potential role of MMP-9/TIMP-1. Methods Diabetic rats were induced with streptozotocin (STZ). Then all rats were maintained for 6 weeks. Routine pathological examination and immnnohistochemistry were made to reveal the histological and cytological appearances. RT-PCR and Western blotting were used to detect the expression of mRNA and protein of MMP-9 and TIMP-1 in the skin. Results Six weeks after STZ treatment, examination of HE-stained skin sections from normal and diabetic animals revealed that the epidermis and dermis layers were thinner in diabetic rats than those in control rats. The skin of diabetic rats showed features of atrophy such as disorganization of connective tissue fiber bundles and enlarged space between collagen fiber bundles. In contrast, thick bundles of connective tissue were observed in the dermis of normal rat skin. In normal skin, cells had a bipolar, spindle-shaped appearance in the thick collagen bundles, while in the skin of diabetic animals the interstitial cells had a rounded, shrunken and crenated appearance. The relative values of expression of MMP-9 in diabetic group were higher than those in normal group with significant difference, however, the relative values of expression of TIMP-I in diabetic group were lower than those in control group. Conclusion The changes in cutaneous histology and cytology appear earlier than skin wound. These "underlying disorders" may be associated with the imbalance of MMP-9/TIMP-1.

Quantification of thermal damage in skin tissue / 西安交通大学学报·英文版

Skin thermal damage or skin burns are the most commonly encountered type of trauma in civilian and military communities. Besides, advances in laser, microwave and similar technologies have led to recent developments of thermal treatments for disease and damage involving skin tissue, where the objective is to induce thermal damage precisely within targeted tissue structures but without affecting the surrounding, healthy tissue. Further, extended pain sensation induced by thermal damage has also brought great problem for burn patients. Thus, it is of great importance to quantify the thermal damage in skin tissue. In this paper, the available models and experimental methods for quantification of thermal damage in skin tissue are discussed.

In vitro amplification of immortalized keratinocytes with micro-carriers in rotary cell culture system / 第三军医大学学报

Objective To explore the way and technique to gain adequate seed cells for skin tissue engineering sufficiently. Methods Human telomerase reverse transcriptase (hTERT) gene was introduced into rabbit keratinocytes by eukaryotic vector. The positive clones were selected and cultured in microcarrier-RCCS. The growth of immortalized keratinocytes was observed, and the metabolic rate and pan-cytokeratins (AE1/AE3) expression of immortalized keratinocytes in experimental groups were detected and compared with those in the control group. Results The immortalized keratinocytes in the experimental groups grew rapidly and had a high metabolic rate and shorter population doubling time (PD) (P

Reconstruction and histologic characteristics of bilayer skin substitute inoculated of human hair follicle cells / 第三军医大学学报

Objective To study histologic characteristics of bilayer skin substitute reconstructed by cells from human hair follicle and whether the skin appendage can be induced based on the bilayer skin substitute.Methods After composite chitosan bilayer skin substitute was reconstructed with dermal papilla cells and outer root sheath cells or dermal sheath cells and outer root sheath cells,its histologic characteristics was investigated in vitro and after transplanted onto SD albino rats.Results Composite chitosan bilayer skin substitute reconstructed by cells from hair follicle had closely arranged epithelium cells and outstanding cornification;Epithelial cords linked with epidermis could be seen in dermis.However,there was no certain hair follicle-like structure formation either in vitro or in vivo.Conclusion Hair follicle cells are good source for skin substitute reconstruction,but it can not induce skin appendage formation through skin substitute by now.