The CK15,CD29 Positive Cells' Location In Different Types Of Skin And Its Affection In Wound Repairing

PART ONE The distribution of CK15,CD29 positive cells in different types of human skin.Objective:To study the distribution of CK15,CD29 positive cells in different types of human skin.Methods:The skin was divided into four groups according to the appearance of different types of skin.These four groups were scalp with hair,body skin with wolly hair,prepuce without hair and foot palm skin without hair.6 specimens were collected per group and these specimens were from 10-30yr old people without gender restriction.3 paraffin sections were picked to do the HE staining,CK15 and CD29 immunohistochemical staining,and then studied under the microscope.The CK15 and CD29 immunohistochemical staining sections were statistically analyzed with image-pro plus software and stata 8.0 statistical software.Results:In general observation,the follicles in the scalp with hair were bigger and with higher density than the follicles in the body skin with wolly hair,and also the hair in the scalp were rougher and longer than the wolly hair in the body skin;prepuce and foot palm skin without hair didn't have follicles.In HE staining sections,the ridges of the epidermis were rougher and less in the scalp with hair than in the body skin with wolly hair;the sebaceous glands in the scalp with hair were also more than the body skin with wolly hair.Prepuce and foot palm skin without hair didn't have follicles,and the ridges of epidermis in the feet skin were thinner and much more than in the prepuce.In CK15 immunohistochemical staining,the positive cells were mainly at the bulge of the hair follicle outer root sheath(ORS) in the scalp with hair and could not be found at the basal of epidermis between follicles;in the body skin with wolly hair,the positive cells were mainly at the basal of epidermis and the staining at the ridges of epidermal basal was darker than at the epidermis basal which was at the top of the dermal papilla.In the prepuce,they were mainly at the epidermal basal,while in foot palm skin they hardly could be observed.In CD29 immunohistochemical staining,except having the same staining as the CK15 marker,there were some differences:the positive cells could be found at the basal of the epidermis between follicles in CD29 staining in the scalp and they could also be found at the basal of sebaceous glands in all the specimens.The results of statistical analysis showed that CK15,CD29 positive cells in the scalp with hair and the prepuce without hair were rich,while in the foot palm skin they were rare,and the difference between these four groups was statistically significant(P＜0.01).Conclusions:The distribution of skin stem cells that are marked with CK15,CD29 is different:in scalp with hair,the stem cells are mainly at the budge of hair follicle ORS;in body skin with wolly hair,they are mainly at the ridges of epidermal basal;in the prepuce and foot palm skin without hair,they are at the basal of epidermis.The CD29 positive cells at the basal of epidermis between follicles in the scalp and at the basal of sebaceous glands may mainly be the transit amplifying cells.The quantity of the stem cells in different types of skin are different:the scalp and prepuce have the biggest quantity of the stem cells,while in the foot palm skin the quantity of the stem cells is the smallest.PART TWO The distribution of CK15,CD29 positive cells in different types of SD rats skin.Objective:To study the distribution of CK15,CD29 positive cells in different types of SD rats skin.Methods:The skin was divided into three groups according to the appearance of different types of skin,that were rat moustache skin,rat body skin and rat foot palm skin.3 paraffin sections were picked to do the HE staining,CK15 and CD29 immunohistochemical staining,and then studied under the microscope.The same methods as the part one were used to analyze the results.Results:In general observation,we found that the follicles in the moustache skin were bigger and isolating easier than in the body skin and the foot palm skin didn't have follicles.In HE staining,there was only one layer of basal cells in the epidermis of moustache skin,and the spinous cells layer and the granulose cells layer were discontinuous in the body skin epidermis.The rat foot palm skin epidermis was similar as human foot palm skin.In CK15 and CD29 immunohistochemical staining,the positive cells were mainly found at the lower half of the hair follicle ORS and the hair ball in the moustache skin and the body skin,while in the foot palm skin,they were mainly found at the basal of epidermis,and could also be found at other layers of epidermis as well.The statistical analysis showed no difference between these three groups(P＞0.05).Conclusions:The distribution of the skin stem cells that are marked with CK15,CD29 in different types of SD rats skin is different:In the moustache skin and body skin,they mainly exist at the lower half of the hair follicle ORS and the hair ball,while in the foot palm skin they exist at the basal and other layers of epidermis.PART THREE The affection of different types of skin which have the CK15,CD29 positive cells in the rat wound repairing.Objective:To study the affection of CK15,CD29 positive cells which exist in the epidermal basal and hair follicle ORS in the rat wound repairing.Methods:The SD rats' skin was divided into three groups according to the appearance of different types of skin,that were follicle with epidermis,follicle without epidermis and foot palm skin.Two full-thickness skin defect wounds were operated on each side of the rat back.The wound size was 1.5×1.5cm~2.6 follicles or 6 pieces of skin were transplanted on each wound,and the wounds were carefully packed.The dressing was moved one week later.The wound size was measured and the specimens were taken at one week,two week and three week after the operation.The specimens were embedded in paraffin and sliced,and the HE staining,CK15 and CD29 immunohistochemical staining were performed,and then studied under the microscope.The same methods as part one were used to analyze the results.Results:All the grafts were surviving well except a few of them were moved in the dressing one week later,two follicle groups were almost epidermidalized two weeks later, and all groups were completely epidermidalized three weeks later.All wounds shrinkaged week after week.The size of the wounds between three groups had no difference statistically(P＞0.05).In HE staining,we observed that the follicle groups were healed better than the foot palm skin group,the thickness of the newly formed epidermis in the follicle groups was much more consistent than the foot palm skin group,and the glands in the dermis appeared and matured earlier and much more than the feet skin group.The wounds in the two follicle groups were almost the same.However,the glands in the follicle with epidermis group were more than the follicle without epidermis group.In the CK15 and CD29 staining,the positive cells could be found in all layers of the newly formed epidermis in all groups.With the extension of time we didn't observed the differences between three groups(P＞0.05).Conclusions:The affection of follicle is better than the affection of foot palm skin in wound repairing.The ability of forming glands in the follicle transplanted wounds is much better than the foot palm skin transplanted wounds.This result demonstrated that the differentiation ability of the stem cells at follicles is better than the stem cells at the epidermal basal.That the quantity of stem cells in all the wounds groups has no difference shows that the different grafts have no effect on the quantity of the stem cells in the newly formed epidermis.