Artificial nerve graft constructed by coculture of activated Schwann cells and human hair keratin for repair of peripheral nerve defects

Studies have shown that human hair keratin(HHK) has no antigenicity and excellent mechanical properties. Schwann cells, as unique glial cells in the peripheral nervous system, can be induced by interleukin-1β to secrete nerve growth factor, which promotes neural regeneration. Therefore, HHK with Schwann cells may be a more effective approach to repair nerve defects than HHK without Schwann cells. In this study, we established an artificial nerve graft by loading an HHK skeleton with activated Schwann cells. We found that the longitudinal HHK microfilament structure provided adhesion medium, space and direction for Schwann cells, and promoted Schwann cell growth and nerve fiber regeneration. In addition, interleukin-1β not only activates Schwann cells, but also strengthens their activity and increases the expression of nerve growth factors. Activated Schwann cells activate macrophages, and activated macrophages secrete interleukin-1β, which maintains the activity of Schwann cells. Thus, a beneficial cycle forms and promotes nerve repair. Furthermore, our studies have found that the newly constructed artificial nerve graft promotes the improvements in nerve conduction function and motor function in rats with sciatic nerve injury, and increases the expression of nerve injury repair factors fibroblast growth factor 2 and human transforming growth factor B receptor 2. These findings suggest that this artificial nerve graft effectively repairs peripheral nerve injury.

Human hair follicle-derived mesenchymal stem cells:Isolation,expansion,and differentiation

Hair follicles are easily accessible skin appendages that protect against cold and potential injuries.Hair follicles contain various pools of stem cells,such as epithelial,melanocyte,and mesenchymal stem cells(MSCs)that continuously self-renew,differentiate,regulate hair growth,and maintain skin homeostasis.Recently,MSCs derived from the dermal papilla or dermal sheath of the human hair follicle have received attention because of their accessibility and broad differentiation potential.In this review,we describe the applications of human hair follicle-derived MSCs(hHF-MSCs)in tissue engineering and regenerative medicine.We have described protocols for isolating hHF-MSCs from human hair follicles and their culture condition in detail.We also summarize strategies for maintaining hHF-MSCs in a highly proliferative but undifferentiated state after repeated in vitro passages,including supplementation of growth factors,3D suspension culture technology,and 3D aggregates of MSCs.In addition,we report the potential of hHF-MSCs in obtaining induced smooth muscle cells and tissue-engineered blood vessels,regenerated hair follicles,induced red blood cells,and induced pluripotent stem cells.In summary,the abundance,convenient accessibility,and broad differentiation potential make hHF-MSCs an ideal seed cell source of regenerative medical and cell therapy.

FT-IR Microspectroscopic Imaging of Cross-Sectioned Human Hair during a Bleaching Process

We investigated bleached human hair by FT-IR microspectroscopy and chemical imaging. The cross sectioned hair is approximately 90 μm in diameter, showed cuticle and cortex in chemical imaging. Differential amide I/II absorbance ratio and broadening amide I band between in the cortex and cuticle were confirmed in FT-IR microspectroscopy and chemical imaging. The cystine monoxide band from the products of disulfide oxidation of the amino acid cystine is associated with hair damaging during bleaching process. With increase bleaching time, the band for cystine monoxide shows more intense and larger area in chemical image. The spatially chemical change was investigated in detail by FT-IR microspectroscopy and chemical imaging during the bleached process.

Ultrastructural analysis shows persistence of adhesion and tight junction proteins in mature human hair

The differentiation of cells composing mature human hairs produces layers with different corneous characteristics that would tend to flake away one from another,as in the corneous layer of the epidermis,without anchoring junctions.It is likely that cell junctions established in the forming cells of the hair bulb are not completely degraded like in the corneous layer of the epidermis but instead remain in the hair shaft to bind mature cuticle,cortex,and medulla cells into a compact hair shaft.During cell differentiation in hairs,cell junctions seem to disappear,and little is known about the fate of junctional proteins present in the mature human hair shaft.The present ultrastructural immunogold study has detected some marker proteins of adhesion junction(cadherin and beta-catenin)and tight junctions(occludin and cingulin)that are still present in cornified hairs where numerous isopeptide bonds are detected,especially in the medulla.This qualitative ultrastructural study indicates that aside from the cell membrane complex,a long corneo-desmosome bonding cortex and cuticle cells,also sparse adherens and tight junction remnants are present.It is suggested that the cornification of these junctions with the incorporation of their proteins within the mature corneous material of the hair shaft likely contributes to maintaining the integrity of the mature hair.This information will also allow us to evaluate the effects of different chemical components present in hair formulations and stains on these junctional proteins and the consequent integrity of the hair shaft.

Bleaching of Black Human Hair

Bleaching of black human hair has been studied systematically. On the basis of experimental data the technology of human hair bleaching through five processes was established. The optimum technology of improving the whiteness and reducing damage on fibers has been found. The technology can provide good luster,smooth handle and relatively high strength retention to human hair used for wigs or drama articles, meeting the needs of people better. Moreover, it also has important reference value to bleaching of other colored fibers.

Analysis of Heavy Metals in Human Hair Using Atomic Absorption Spectrometry (AAS)

Hair samples of individual were analysed for heavy metals (Cd, Cr, Pb and As) across gender and various occupational distributions by Atomic Absorption Spectrophotometric technique (AAS). The results of replicate analysis shows the following mean concentrations (mg/kg): Cd = 27.8 ± 8.0, Cr = 2.70 ± 0.7, Pb = 73.8 ± 42.3 and As = 222 ± 34.1. The coefficients of variation for the total distribution is;Cd = 28%, Cr = 26%, Pb = 57% and As = 15%. The distribution of the metals follows the series in decreasing order of As > Pb > Cd > Cr, while their coefficients of variation is in the order of Pb > Cd >Cr > As. The result shows the presence of all the metals in relatively large amounts with As having the highest concentration between the two genders. The difference between male and female concentration could be due to individual differences in exposure to heavy metal load as a result of habitual or environmental factors.

PREPARATION AND CERTIFICATION OF THE HUMAN HAIR REFERENCE MATERIAL

The preparation of the human hair reference material and the steps taken to confirm its homogeneity and stability as well as its certificatipn are described. Certified values for 17 elements, which are of importance for human health, and reference values for the other 13 elements are provided for this material.

DISTRIBUTION OF SOME ELEMENTS IN HUMAN HAIR AND ORGANS DETERMINED BY NAA

Twenty four male autopsies were obtained from Shanghai. The samples of liver, lung, kidney cortex, brain and scalp hair were collected from the autopsies. The elements As, Br, Ca, Cd, Cl, Co, Cr, Cu, Fe, Hg, Mg, Mn, Na, Rb, 8, Se and Zn were determined by neutron activation analysis (NAA). The present study provides the elemental levels of different tissues for normal male adults. The distribution pattern of elements was discussed. Great difference on distribution of some elements in different tissues was observed.

Degradation of human hair keratin scaffold material used to repair injured skeletal muscles of rabbits

Objective:To explore the mechanism of the degradation of human hair keratin (HHK) scaffold material implanted in damaged skeletal muscle tissues. Methods: Six New Zealand rabbits with HHK scaffold material implants (composed of 3 different types of HHK material with varied degradation speed) after musclectomy were divided into 3 groups (2 in each group) to observe the degradation of the material at 1, 3, 6weeks after operation. Another rabbit without operation was used as the control group. The degradation of HHK was observed with light microscopy, histochemistry of ubiquitin and electron microscopy. Results:Light microscopy showed that human hair cuticles fell off from the HHK material and emerged, and the macrophagocytes and multinucleate giant cells were attached onto the surface of the material, which became homogeneous at the first postoperative week. The HHK scaffold material was degraded into particles that was phagocytosed by macrophagocytes and multinucleate giant cells at the third week. Ubiquitin enzymatic histochemistry showed that the macrophagocytes and the multinucleate giant cells were positive at the first week. Under electron microscope, HHK scaffold material was degraded into particles, and at the sixth week,part of HHK scaffold material was further degraded. Conclusion: Large mass of the HHK scaffold material is degraded via ubiquitin system, and the resultant particles are phagocytosed and degraded with the cooperation of lysosome and ubiquitin.

Peripheral nerve repair by conduits made of human hair keratin: An experimental study

Objective: To explore the method to repair injured peripheral nerve using conduits made of human hair keratin (HHK). Methods: The tibial nerves of rabbits were transected leaving a gap 10 mm in length between the 2 severed ends, which were either routinely sutured or bridged using HHK nerve conduits. Electro-physiological , anatomical and histological examinations were performed at different time postoperatively. Results: Electrophysiological study showed more obvious improvement in the neural function recovery in rabbits with HHK conduits bridging as compared with that in rabbits with routine suture. In the former group, HHK conduits were gradually degraded and absorbed with large amount of myelinated nerve fibers and Schwann cells regenerated around HHK conduits. In the latter group, however, the nerve tissues around the suture were degenerated and replaced by connective tissues. Conclusion: HHK may induce the regeneration of the nerve fibers and provides an ideal approach to repair nerve damages.

Reduction and Block Staining of Human Hair Shafts and Insect Cuticles by Ammonium Thioglycolate to Enhance Transmission Electron Microscopic Observations

We have developed a method where, after glutaraldehyde fixation, human hair shafts and insect cuticles are treated with ammonium thioglycolate (ATG) to improve ultrastructural staining. Conventional transmission electron microscopic (TEM) preparations do not distinguish the A-layer and the exocuticles of hair shafts. However, after ATG treatment, the A-layer appears in higher contrast. ATG treatment has also been used to observe the fibrillar structure in the cortex. In the cuticle of beetles, the epicuticle is stained by ATG. Although the human hair shaft (keratin) and insect cuticle (chitin) are composed of different materials, both can be reduced by the ATG solution. The ammonium in the ATG solution reacts with hair shafts and insect cuticles, causing a reduction of swelled cuticles. Therefore, ATG not only stains, but also reduces human hair shafts and the cuticles of beetles.

Human hair follicle-derived mesenchymal stem cells promote tendon repair in a rabbit Achilles tendinopathy model

Background:Hair follicles are easily accessible and contain stem cells with different developmental origins,including mesenchymal stem cells(MSCs),that consequently reveal the potential of human hair follicle(hHF)-derived MSCs in repair and regeneration.However,the role of hHF-MSCs in Achilles tendinopathy(AT)remains unclear.The present study investigated the effects of hHF-MSCs on Achilles tendon repair in rabbits.Methods:First,we extracted and characterized hHF-MSCs.Then,a rabbit tendinopathy model was constructed to analyze the ability of hHF-MSCs to promote repair in vivo.Anatomical observation and pathological and biomechanical analyses were performed to determine the effect of hHF-MSCs on AT,and quantitative real-time polymerase chain reaction,enzyme-linked immunosorbent assay,and immunohistochemical staining were performed to explore the molecular mechanisms through which hHF-MSCs affects AT.Furthermore,statistical analyses were performed using independent sample t test,one-way analysis of variance(ANOVA),and one-way repeated measures multivariate ANOVA as appropriate.Results:Flow cytometry,a trilineage-induced differentiation test,confirmed that hHF-derived stem cells were derived from MSCs.The effect of hHF-MSCs on AT revealed that the Achilles tendon was anatomically healthy,as well as the maximum load carried by the Achilles tendon and hydroxyproline proteomic levels were increased.Moreover,collagen I and III were upregulated in rabbit AT treated with hHF-MSCs(compared with AT group;P<0.05).Analysis of the molecular mechanisms revealed that hHF-MSCs promoted collagen fiber regeneration,possibly through Tenascin-C(TNC)upregulation and matrix metalloproteinase(MMP)-9 downregulation.Conclusions:hHF-MSCs can be a treatment modality to promote AT repair in rabbits by upregulating collagen I and III.Further analysis revealed that treatment of AT using hHF-MSCs promoted the regeneration of collagen fiber,possibly because of upregulation of TNC and downregulation of MMP-9,thus suggesting that hHF-MSCs are more promising for AT.

Mercury in soil,vegetable and human hair in a typical mining area in China:Implication for human exposure

Concentrations of total mercury（T-Hg） and methylmercury（MeHg） in soil, vegetables, and human hair were measured in a mercury mining area in central China. T-Hg and MeHg concentrations in soil ranged from 1.53 to 1054.97 mg/kg and 0.88 to 46.52 μg/kg, respectively.T-Hg concentrations was correlated with total organic carbon（TOC） content（R^2= 0.50, p〈0.01）and pH values（R^2= 0.21, p〈0.05）. A significant linear relationship was observed between MeHg concentrations and the abundance of sulfate-reducing bacteria（SRB）（R^2= 0.39, p〈0.05） in soil.Soil incubation experiments amended with specific microbial stimulants and inhibitors showed that Hg methylation was derived from SRB activity. T-Hg and MeHg concentrations in vegetables were 24.79 – 781.02 μg/kg and 0.01 – 0.18 μg/kg, respectively; levels in the edible parts were significantly higher than in the roots（T-Hg： p〈0.05; MeHg： p〈0.01）. Hg species concentrations in rhizosphere soil were positively correlated to those in vegetables（p〈0.01）, indicating that soil was an important source of Hg in vegetables. Risk assessment indicated that the consumption of vegetables could result in higher probable daily intake（PDI） of T-Hg than the provisional tolerable daily intake（PTDI） for both adults and children. In contrast, the PDI of MeHg was lower than the reference dose. T-Hg and MeHg concentrations in hair samples ranged from 1.57 to 12.61 mg/kg and 0.04 to 0.94 mg/kg, respectively, and MeHg concentration in hair positively related to PDI of MeHg via vegetable consumption（R^2= 0.39, p〈0.05）, suggesting that vegetable may pose health risk to local residents.

Identification of adrafinil and its main metabolite modafinil in human hair.Self-administration study and interpretation of an authentic case

For several years,the misuse of stimulant substances is increasingly observed both in the field of sport,to improve the functions of the body and therefore to be more performant,and also by non-athletes to make life more tolerable on a daily basis.Adrafinil,2-((diphenylmethyl)sulfinyl)-N-hydroxyacetamide,is a drug designed for the treatment of narcolepsy by promoting an awakened state,and to treat alertness and neurological symptoms in the elderly.It is primarily metabolized in vivo to an active form,i.e.modafinil,2-((diphenylmethyl)sulfinyl)acetamide.The World Anti-Doping Agency(WADA)banned these two drugs in sports in 2004.The authors report an authentic case involving adrafinil and modafinil.The laboratory was requested to test for adrafinil in a hair strand collected from a woman found in posses-sion of vials of adrafinil and suspected of trafficking.A specific method was developed by liquid chromatography tandem mass spectrometry(LC-MS/MS).Unlike modafinil(varying from 6.8 to 13.9ng/mg),adrafinil was not identified in the strand.The interpretation of the results was difficult because this is the first case describing human hair analysis.In order to be able to interpret the results,a self-administration study was conducted after an oral administration to a volunteer(200 mg)whose beard hair was collected 10days after adminis-tration.The analysis of this specimen highlighted the presence of adrafinil at 0.8ng/mg and modafinil at 0.5 ng/mg.These results demonstrate the dual identification of both compounds after a single consumption,even after administration of a low dose.According to these results,the analysis of the hair strand from the authentic case does not match with a con-sumption of adrafinil,in accordance with abuse of modafinil alone.Intelligence considered that this was a trafficking case of adrafinil,with no self-consumption.

Flavonoids in safflower extract reduce cisplatin-induced damage to human follicle dermal papilla cells by inhibiting DNA damage and Rad17/Chk1/Cdc25C signaling

Background:Cisplatin is a chemotherapeutic agent commonly used clinically for the treatment of various human cancers.Patients often reduce the use of cisplatin due to its side effects,which in turn affects its treatment.This study explored the mechanism of action of safflower extract as an adjuvant traditional Chinese medicine for chemotherapy.Methods:Primary human follicle dermal papilla cells(HFDPCs)were used as target cells for cisplatininduced damage to hair cells.Western blotting was used to investigate the molecular targets of cisplatin and safflower extract in causing HFDPCs damage.Cell survival and cell cycle were analyzed by mitochondrial staining reagent WST-1 and propidium iodide.Results:Cisplatin could reduce the viability of HFDPCs without causing cell death.Cisplatin increased the level of phospho-Rad17 in HFDPCs and activated the Chk1/Cdc25C signaling to reduce the expression of Cdc2 protein,thereby arresting the cells in the G2/M phase.The combination of safflower extract and the flavonoids could effectively inhibit the signal transduction of Rad17/Chk1/Cdc25 in cisplatin-treated cells and reduce the cell population in the G2/M phase.Finally,we also confirmed that safflower extract could effectively inhibit the damage to HFDPCs caused by cisplatin,mainly at the level of reducing the DNA damage caused by cisplatin.Conclusions:Safflower extract can be used as an adjuvant Chinese medicine for chemotherapy to reduce the damage caused by chemotherapy to normal hair follicle cells.

Analysis of Heavy Metals in Human Scalp Hair Using Energy Dispersive X-Ray Fluorescence Technique

Analysis of six heavy metals (Cr, Mn, Ni, Cu, Zn and Mo) in human scalp hair was carried out among various occupational distributions to ascertain their heavy metal burden, using energy dispersive x-ray fluorescence technique (ED-XRF). The result of the analysis shows that mean concentrations (mg/kg) of heavy metals obtained were as follows: Cr = 17.1 ± 12.7;Mn = 3.11 ± 0.50;Ni = 11.3 ± 9.3;Zn = 451 ± 128;Cu = 83.3 ± 35.8 and Mo is 9.16 ± 9.1.While the mean concentrations of Cr, Cu, and Mo were higher in the females, that of Mn, Ni and Zn were more in the males. Statistical analysis of the results for both genders at 0.05 probably shows significant difference for Ni, Zn and Mo while Cr, Mn and Cu showed no significant difference. The relationships between age, body mass, height, and heavy metal concentrations were also investigated. Statistical analysis of the results indicates that there was no correlation between the body mass (R2 ≤ 0.048), height (R2 ≤ 0.002) and heavy metal concentration in hair. Zn showed the highest deviation among other elements in the individual samples for both genders which reflect the individual variation in the concentration of Zn.

TOTAL AND METHYL MERCURY LEVELS IN HUMAN SCALP HAIRS

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Therapeutic Approach for Hair Growth and Regeneration Using Bioactive Formulation Containing Mesenchymal Stromal Cell-Derived Conditioned Medium

Background and Aims: Androgenetic alopecia (AGA) is a common form of hair loss in both men and women. Despite its high prevalence and associated patient morbidity, the approved therapeutic options are limited to finasteride and minoxidil. The present study is aimed at assessing the efficacy of hair serum formulation, Trichosera®containing Bone marrow-derived mesenchymal stromal cells conditioned media as an active ingredient, for hair fall control and hair regrowth in healthy Indian human volunteers. Methods: The product was made using a 20% concentration of 10X Conditioned Media along with excipients. The final product was tested for physicochemical parameters, biomarkers, total protein content and microbial limits as per our in-house specifications. Results: The primary irritation patch test showed that the product is non-irritant and dermatologically safe. A clinical study on 40 subjects was conducted to evaluate the effectiveness of the bioactive formulation in hair fall control and hair regrowth in healthy volunteers. Phototrichogram measurement showed hair density and hair growth rate increased significantly by 11.54% and 18.66% at week 24. Hair tensile strength also increased significantly by 41.10% at 12 weeks follow-up. Hair pull test, to see a reduction in pulled hair and comb’s test to show a decrease in hair fall significantly improved from week 4 onwards. There were no significant adverse events in response to the product application. Conclusion: It is concluded that the hair serum product is completely safe on direct application to the scalp and showed significant improvement in the hair growth rate, hair density, scalp condition and reduction in hair fall. .

人毛囊真皮鞘细胞移植治疗兔跟腱炎实验研究

目的:探讨人毛囊真皮鞘细胞(hHF-DSCs)移植对兔跟腱炎的治疗效果。方法:采集人后枕部毛囊单位,并进行分离培养获得hHF-DSCs。选取30只雄性大白兔随机分为对照组、模型组和治疗组,每组10只。模型组及治疗组于兔左侧跟腱止点上1 cm处多点注射Ⅰ型胶原酶建立跟腱炎兔模型。建立跟腱炎模型成功后于第0、3、6、13天,治疗组在兔跟腱患侧注射0.5 ml的hHF-DSCs,模型组与对照组在兔跟腱相同部位注射相同体积0.9%氯化钠溶液。观察各组兔跟腱外观以及组织病理学染色结果。采用ELISA法检测跟腱组织羟脯氨酸(HYP)含量以及肌腱蛋白C(TNC)、基质金属蛋白酶-9(MMP-9)、白细胞介素(IL)-1β、IL-6、IL-10和肿瘤坏死因子-α(TNF-α)表达水平。采用免疫组化法检测跟腱组织Ⅰ、Ⅲ型胶原(collagenⅠ、Ⅲ)表达水平。采用牵拉法检测跟腱组织生物力学。结果:分离培养后的hHF-DSCs表面CD14、CD34、CD45和人类白细胞抗原-DR(HLA-DR)几乎不表达,CD49d、CD90、CD73和CD105表达率接近100%。与对照组比较,模型组步态改变明显,跟腱红肿、无弹性、无光泽。HE染色结果显示,模型组胶原纤维排列疏松,原核细胞较少,分布紊乱不均匀,表明跟腱炎兔模型构建成功。治疗组跟腱组织弹性和光泽度好于模型组,且治疗组HE染色显示胶原纤维排列整齐,无明显纤维坏死,炎症细胞较少,胶原纤维原核细胞数量正常。模型组HYP含量低于对照组和治疗组,对照组高于治疗组(均P<0.05)。模型组MMP-9水平高于对照组和治疗组,治疗组高于对照组(均P<0.05)。模型组TNC水平低于对照组和治疗组,治疗组低于对照组(均P<0.05)。与对照组比较,治疗组和模型组兔跟腱组织IL-1β、IL-6、IL-10及TNF-α表达水平升高(均P<0.05)。与模型组比较,治疗组IL-1β、IL-6、TNF-α表达水平降低(均P<0.05)。与模型组比较,治疗组collagenⅠ、Ⅲ升高(均P<0.05)。与对照组比较,模型组collagenⅠ、Ⅲ降低(均P<0.05)。模型组跟腱最大负荷低于对照组,治疗组高于模型组(均P<0.05)。结论:hHF-DSCs移植治疗兔跟腱炎,能够促进受损跟腱组织修复及胶原合成,降低胶原降解,减轻炎症反应。

交直流输电线路混合电场的人体感受试验及设计控制值研究

交直流输电线路同走廊和同塔架设是随着我国电网技术发展而出现的新输电方式,国内外尚无相应的交直流混合电场设计控制值标准。因此,设计同走廊和同塔架设的交直流输电线路时,导线对地高度和走廊宽度的选取面临无据可依的难题。该文提出确定交直流混合电场设计控制值的原则;开展不同交流(工频)电场和直流电场组合下的人体直接感受和暂态电击感受试验;通过对试验结果的统计分析,获得不同概率下人体不出现明显感受时的交流(工频)电场和直流电场关系曲线;在此基础上,得出确定交直流输电线路混合电场设计控制值的公式。该研究可为同走廊和同塔架设交直流输电线路设计中交直流混合电场的控制提供一定技术支撑。