Comparison of epidermal morphologic response to commercial antiwrinkle agents in the hairless mouse.

BACKGROUND: A large number of commercial antiwrinkle and antiaging compounds are available to consumers for rejuvenation of facial skin ravaged by age or solar radiation. Experimental data on the histological effects of these commercial products in laboratory models are sparse. OBJECTIVES: To compare the efficacy of topical application of five commercially available antiaging compounds (retinoic acid, glycolic acid, vitamin C, estrogen, and soy) on the dorsal skin. METHODS AND MATERIALS: The effects were examined using light microscopic analysis of the epidermis in the normal nonirradiated hairless mouse. The agents were applied daily to dorsal tattooed areas for 2 weeks before histological assessment; neighboring untreated surface areas were used as control. Morphometric measurements of total epidermal width, nuclear volume of keratinocytes in three layers, and index of proliferating cell nuclear antigen according to immunohistochemistry were obtained and statistically analyzed. RESULTS: Significant histomorphometric effects were noticed with all five agents, but more pronounced changes were obtained with glycolic acid, estrogen, and retinoic acid product. CONCLUSIONS: These baseline data will be useful for future studies on the effect of ultraviolet radiation to cause photoaging and reparative effects of similar agents in this animal. The information contained in the report may provide guidelines to consumers and clinicians.

Association of Diet With Skin Histological Features in UV-B-Exposed Mice.

IMPORTANCE: Long-term exposure to solar radiation produces deleterious photoaging of the skin. It is not known if diet can influence skin photoaging. OBJECTIVES: To study the influence of a calorie-restricted diet and an obesity diet in mice exposed to long-term UV-B irradiation to assess if there is an association between diet and histopathological response to UV-B irradiation. DESIGN, SETTING, AND PARTICIPANTS: In this animal model study in an academic setting, the dorsal skin of SKH1 hairless mice receiving normal, calorie-restricted, and obesity diets was exposed to UV-B irradiation 3 times a week for 10 weeks and were compared with corresponding controls. The mice were placed in the following groups, with 8 animals in each group: (1) intact control (C) with regular diet and no UV-B exposure, (2) intact control with UV-B exposure (CR), (3) calorie-restricted diet (CrC), (4) calorie-restricted diet with UV-B exposure (CrR), (5) obesity diet (OC), and (6) obesity diet with UV-B exposure (OR). The experiment was conducted during October through December 2013. Tissue processing and histological analysis were completed in 2016. MAIN OUTCOMES AND MEASURES: Histomorphometric analysis was performed on paraffin-embedded skin sections stained by histological and immunohistochemical methods for estimation of epidermal thickness, epidermal proliferating cell nuclear antigen index, collagen I, elastic fibers, fibroblasts, mast cells, dermal cellularity, and adipose layer ratio. Changes in wrinkles were noted. RESULTS: Hairless female mice (age range, 6-8 weeks) were obtained. With a normal diet, changes from UV-B irradiation occurred in epidermal thickness, epidermal proliferating cell nuclear antigen index, collagen I, elastic fibers, fibroblasts, and mast cells, which were modestly influenced by an obesity diet. Calorie restriction influenced the skin in nonirradiated control animals, with higher values for most variables. After UV-B exposure in animals with calorie restriction, epidermal thickness was increased, but other variables were unaffected. Animals receiving the calorie-restricted diet lost weight when exposed to long-term UV-B irradiation. Wrinkles were reduced in the calorie-restricted control group and in UV-B-exposed animals who received the obesity diet. CONCLUSIONS AND RELEVANCE: Dietary alterations seem to modify histopathological responses to UV-B exposure in the skin of hairless mice. LEVEL OF EVIDENCE: NA.

Modulation of cutaneous aging with calorie restriction in Fischer 344 rats: a histological study.

OBJECTIVE: To examine whether histological changes in skin owing to intrinsic aging in a laboratory rodent model are modulated by caloric restriction (CR). METHODS: The abdominal skin from colony-raised ad libitum-fed Fischer 344 rats and age-matched rats subjected to CR was studied in the light microscope using histological morphometric methods. Animals 4, 12, and 24 months or older were used in this study. We studied the skin to obtain (1) quantitative data on the depth of the epidermis, dermis, and fat layer, the epidermal cellular density, the percentage fraction of dermal collagen, elastic fibers, pilosebaceous units, and capillaries, and the fibroblast density; and (2) qualitative assessment of histological staining for dermal glycosaminoglycans. We analyzed data by means of general linear model 2-way analysis of variance to obtain significance for the effects of age, diet, and age-diet interaction. RESULTS: The ad libitum-fed rats showed age-related increase in the depth of the epidermis, dermis, and fat layer. Calorie restriction prevented these changes, but epidermal nuclear density appeared to be stimulated. A trend toward increased values for collagen and elastic fibers, fibroblasts, and capillaries in skin samples from CR rats was observed. Pilosebaceous units were not modified. Moderately reduced staining for the dermal glycosaminoglycans in the skin of CR rats was noticed. CONCLUSIONS: Histomorphological changes resulting from intrinsic aging affected some of the studied variables in the rat skin, and these changes were delayed or prevented by CR. Some stimulatory effects, such as increased densities of fibroblasts and capillary profiles and higher values of connective tissue fibers resulting from CR, were also observed. Cutaneous morphological changes due to natural aging in this rat model seem to be modified by physiological or metabolic alterations imposed by CR.

Fibrin tissue adhesive and autologous concha cartilage for reconstruction of the posterior-superior canal wall of the chinchilla middle ear.

HYPOTHESIS: We conducted this study to prove that fibrin tissue adhesive (FTA) is safe, efficacious, biocompatible, and readily biodegradable with no deleterious side effects for fixation of a cartilage graft to bone along the chinchilla canal wall. METHODS: A posterior-superior canal defect was created in 12 chinchillas. The canal walls of six chinchillas were closed with autologous concha cartilage alone, whereas the canal wall of the remaining six animals were closed with cartilage in conjunction with fibrin tissue adhesive. RESULTS: Animals were killed 8 weeks postoperatively. Three of six cartilage grafts were displaced in the graft alone group, whereas all six grafts in the cartilage with FTA group healed without displacement. CONCLUSION: Fibrin tissue adhesive was found to be effective, biocompatible, biodegradable, and without any deleterious side effects for reconstruction of the superior-posterior canal wall of chinchillas.

Histomorphologic changes in aging skin: observations in the CBA mouse model.

BACKGROUND: Aging of human skin is a phenomenon resulting from a combination of chronological aging and environmental stressors such as sunlight. OBJECTIVES: To study the effects of intrinsic aging on the skin in laboratory-raised CBA mice in 3 age groups, and to assess histological alterations as a function of age in this model. METHODS: Skin samples from CBA mice in 3 age groups (young, young adult, and old) were obtained from the dorsal and ventral areas, pinna, and hind foot to study the following variables using light microscopic manual morphometric methods: the depth of the epidermis and number of epidermal cells, depth of the dermis, and percentage area of dermal collagen, elastic fibers, pilosebaceous units, blood vessels, and tissue space. The obtained values were analyzed using 1-way analysis of variance to detect any significant effects of age. RESULTS: There was a notable attrition of the epidermal thickness and number of cells that could be correlated with age. A reduced number of pilosebaceous units was noted in skin samples from the dorsal region and the footpad. No conspicuous change was noted in the depth of the dermis or percentage area of collagen in aging animals. A proliferation of stainable elastic fibers was demonstrated in the dorsal skin and footpad of older mice. CONCLUSIONS: CBA mice show unique age-related histological modifications of the skin that are different from other rodent species. These baseline data will be helpful in further studies of regenerative effects of pharmaceutical agents on the histological structure of skin and in photoaging studies.

Cosmeceutical effect on skin surface profiles and epidermis in UV-B-irradiated mice.

IMPORTANCE: These data may be useful for developing guidelines for clinicians and the general population related to the reversal of photoaging effects on the aging face damaged by solar radiation. OBJECTIVE: To investigate antiaging effects of 4 commercially available topical agents on the dorsal skin in photoaged hairless mice. DESIGN AND SETTING: Animal study at an academic medical center. Animals comprised 56 female Skh-1 hairless mice (6-8 weeks old). Skin samples were collected from nonirradiated intact mice (control), mice irradiated with UV-B for 8 weeks, mice irradiated with UV-B and then exposed to a topical cosmeceutical applied for 5 weeks, and UV-B-irradiated mice not exposed to cosmeceuticals and retained for 5 weeks until the end of the experiment. INTERVENTION: The mice were exposed to UV-B light 3 times a week for 2 months, followed by topical application of a peptide, antioxidant, estrogen, and retinoic acid agent for 5 weeks. MAIN OUTCOMES AND MEASURES: Surface features such as wrinkling were analyzed from replicas along with histomorphometric determination of epidermal thickness, sebocyte counts, and immunohistochemical study of proliferating cell nuclear antigen (PCNA). RESULTS: Exposure to UV-B induced significant wrinkle formation after 13 weeks, which was attenuated with treatments with a peptide cream, antioxidant mixture, and estrogen cream (mean [SD] Rz values: control [C], 60.7 [19.0]; irradiated [RAD], 51.8 [15.9] [P < .001]; irradiated-long [RAD-long], 86.0 [28.3] [P = .01]; antioxidant [AO], 45.2 [13.2]; peptide, 63.4 [18.8], estrogen, 64.6 [21.2]; retinoic acid [RA], 73.9 [28.5]; RAD-long vs C [P = .01], vs RAD [P < .001], vs estrogen [P = .04], vs peptide [P = .02], vs AO [P<.001], vs RA [P = .25]. There was a trend of reversal of irradiation-induced augmentation of epidermal thickness in animals treated with the peptide and AO (mean [SD] epidermal width: C, 21.0 [2.2] µm; RAD, 41.3 [7.0] µm [P < .001]; RAD-long, 39.1 [11.0] µm [P = .006]; AO, 37.3 [14] µm [P < .001]; peptide, 33.9 [3.8] µm [P = .01]; estrogen, 59.2 [9.2] µm [P = .003]; RA, 52.4 [8.7] µm [P < .001]). Retinoic acid augmented epidermal width and sebocyte counts (mean [SD] sebocyte data [number per gland]: C, 9.4 [2.0]; RAD, 11.69 [1.5] [P < .001]; RAD-long, 6.5 [1.3] [P = .73]; peptide, 7.2 [1.7] [P = .03]; estrogen, 4.1 [0.9] [P < .001]; AO, 7.2 [1.7] [P = .06]; RA, 11.0 [1.4] [P = .01]). Estrogen cream was effective in restoring surface features but enhanced thickness of epidermis in irradiated specimens. All groups had a higher PCNA index score except for peptide treatment, which brought it down to the control level (mean [SD] PCNA index values: C, 17.3 [1.5]; RAD, 32.4 [6.8] [P < .001]; RAD-long, 34.0 [6.1] [P < .001]; AO, 62.1 [3.5] [P = .01]; peptide, 20.1 [6.3] [P < .001]; estrogen, 56.8 [10.0] [P < .001]; RA, 35.2 [10.2] [P < .001]). CONCLUSIONS AND RELEVANCE: Of the 4 cosmeceuticals tested within this experimental period, peptide cream and antioxidant mixture were the most effective overall in reversing photoaging effects; retinoic acid was the least effective of these topical agents. LEVEL OF EVIDENCE: NA.

Effects of topicals on the aging skin process.

This article summarizes the antiaging properties of retinoids, glycolic acid, ascorbic acid, and peptide topicals. The supporting evidence is taken from the literature and the primary author's research, consisting of previously published data and new results from ongoing projects.

Profilometric and morphometric response of murine skin to cosmeceutical agents.

OBJECTIVE: To investigate whether topical antiaging compounds can reduce wrinkle depth as noted at replica profilometry with comparable changes in histologic findings in hairless mice. METHODS: Commercial retinoic acid cream, a peptide lotion, and a soy cream were applied to the dorsal skin for 4 weeks. Silicone-negative replicas of treated and untreated skin surface were photographed and evaluated for traditional features of surface roughness. Skin samples were processed using histomorphometry and immunohistochemistry of proliferating cell nuclear antigen. Quantitative light microscopic data were acquired for estimating replication of epidermal keratinocytes, epidermal thickness, and depth of dermal collagen bundles. RESULTS: Data were analyzed by comparing means with 1-way analysis of variance, and significant changes in all measurements were noted. Augmented keratinocyte proliferation and thickening of viable epidermis were observed with all 3 compounds, although a greater effect was found in the retinoic acid and peptide treatment groups. A similar trend was noted with respect to widening of the collagen layer. Epidermal surface roughness manifested maximum smoothing after treatment with the peptide compound. CONCLUSION: The pronounced effects noted with all 3 compounds indicate that topical agents other than retinoic acid may have comparative stimulating effects on the skin in nonirradiated mice.

Vestibular cytotoxicity in gentamicin-treated frogs: a preliminary report.

OBJECTIVE: The aim of this study is to determine the immediate effects of intraperitoneal doses of gentamicin (GM) which would result in variable degrees of destruction of crista ampullary hair cells of frogs. This information will serve as a baseline guide to cell regeneration experiments on the damaged vestibular sense organ. STUDY DESIGN: The American bullfrog was administered daily intraperitoneal doses of 50, 100, 150, and 200 mg/kg of GM for 7 days. Animals were sacrificed 1 day after the final injection for cytomorphic evaluation. Histologically processed posterior semicircular duct cristae were resin-embedded, and tissue samples were subjected to serial cross sectioning of the crista from the periphery to the central zone using glass knives in an ultramicrotome. Stained sections were analyzed in light microscope using ocular grid micrometry. The areal density of nuclear profiles of the vestibular sensory and supporting cells (sensory cells [SNCs] and supporting cells [SPCs], respectively; number per square millimeter) and the nuclear diameter of SNCs were manually determined. RESULTS: A 7-day administration of GM produced noticeable quantitative alteration of the posterior crista hair cells and SPCs. Histological analysis revealed a significant decrease in the density of SNCs and a concomitant increase in the density of SPCs (1-way analysis of variance). CONCLUSION AND SIGNIFICANCE: The cytomorphic data derived from this study show that 4 doses of intraperitoneal gentamicin administered to the bullfrog caused a decline in the areal density of sensory hair cells of the posterior canal crista ampullaris. Also noted was an increase in the density of adjacent SPCs. Although speculative, the increase in SPC population could be a harbinger of regeneration of the vestibular hair cells as suggested by other investigators in different species. The significance of present observations will be helpful to initiate future studies related to recovery of SNCs in a similarly damaged frog ampullary organ. Through a standardized quantitative approach to the study of SNCs and SPCs of the crista organ, the vestibulo-toxicity of newly developed drugs can be assessed.

Histological evaluations following 1,064-nm Nd:YAG laser resurfacing.

BACKGROUND AND OBJECTIVES: The long pulse 1,064-nm Nd:YAG laser is used clinically to decrease rhytid formation. The dermal level at which this change occurs has not been established. This study attempts to answer these questions using a porcine skin model. STUDY DESIGN/MATERIALS AND METHODS: Non-randomized prospective experimental trial involving the domestic piglet treated serially with the long pulse 1,064-nm Nd:YAG laser. RESULTS: Collagen formation occurred at the level of the reticular dermis. After one laser treatment, a significant level of collagen formation was induced in the reticular dermis compared to controls. The greatest gain was observed after four laser treatments. Energy levels of 20, 30, 40, and 50 J/cm2 were evaluated. Although not statistically significant, 30 J/cm2 had the greatest effect on collagen formation. However, at 50 J/cm2, marked ablative changes to the epidermis were observed. CONCLUSIONS: The long pulse 1,064-nm Nd:YAG laser induces collagen formation in the reticular dermis in porcine skin.