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Local Doxycycline,DHA,Vita.D3 Administration Reduces Scarring and Improves Wound Healing by immunomodulation.

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#1 Ruth

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Posted 10 July 2018 - 06:10 AM

PURPOSE: Scars can be aesthetically and functionally devastating. Novel scar treatments have the potential to impact millions of patients. Doxycycline possesses known anti-fibrotic properties. However, its role as a potential vulnerary agent has remained unexplored. We hypothesized that topically applied doxycycline would reduce scarring.

METHODS: We employed a murine wounding model wherein full-thickness wounds are stented with silicone rings. This prevents wound contraction and mimics human wound healing kinetics. Antibiotic and PBS (control) solutions were injected locally into the superficial aspect of the wound base following surgery. Wounds were harvested upon complete re-epithelialization (day 15) for tensile strength testing and histologic examination. To quantify scar thickness, a blinded observer analyzed images of H&E-stained wound cross sections in Adobe Photoshop. Three photos were analyzed per wound; for each, dermal thickness was measured at three different scar depths, for a total of nine measurements per wound. Scar thickness was calculated as the mean of these measurements. To quantify collagen branching, picrosirius red-stained images were analyzed using an algorithm in MATLAB to calculate branchpoints per 100 micron2. Bacterial colonization of wounds was assessed via wound swabbing and culture for 24 hours. Cell migration was assessed using an in vitro scratch assay. The population of “scarring” fibroblasts (Engrailed 1-positive fibroblasts, EPFs) in wounds was determined by using the same methods in En1 Cre ;ROSA26 mTmG mice.

RESULTS: Doxycycline treatment significantly reduces scar dermal thickness by 37% compared to PBS (*P<0.001). Picrosirius red staining illustrates that doxycycline-treated wounds have significantly reduced picro-red, scar-like collagen (*P=0.016), and increased picro-green, favorable collagen (*P=0.016). These picro-green fibers also demonstrate significantly more branching (*P=0.032) and are less aligned, like unwounded skin collagen. Notably, ultimate tensile strength is comparable between doxycycline-treated and PBS-treated wounds (respectively: 0.462 MPa, n=10; 0.534 MPa, n=9; P=0.438). Bacterial colonization is not significantly altered by doxycycline treatment (P>0.05 at days 1, 3, 5, 7, and 9). Other tetracycline antibiotics (specifically, minocycline and tetracycline) do not decrease scar thickness. “Scarring” fibroblasts (EPFs) are reduced in doxycycline-treated wounds, demonstrated by a 33.7% decrease in GFP signal in wounds from En1 Cre ;ROSA26 mTmG treated mice (n=6, *P=0.021). Treating “scarring” fibroblasts (EPFs) with doxycycline in vitro significantly reduces migration rate (n=4, *P<0.001).

CONCLUSIONS: Locally administered doxycycline reduces scarring without sacrificing scar strength. These findings may be due to increased picro-green collagen that mimics unwounded skin collagen. Our results suggest that doxycycline’s effects are not related to antimicrobial activity. Rather, doxycycline may alter scarring fibroblast behavior during healing. Collectively, our data suggest that doxycycline may represent a novel anti-scarring therapy with the benefits of a well-established safety and dosing profile. We favor rapid transition to studies in human patients to determine whether similar effects are observed.



Wound healing is a physiological event that generates reconstitution and restoration of granulation tissue that ends with scar formation. As omega fatty acids are part of membrane phospholipids and participate in the inflammatory response, we investigated the effects of omega-3, omega-6, and omega-9 fatty acids in the form of oils on wound healing. Linseed (LO), evening primrose (EPO), and olive oils (OO) rich in omega-3, omega-6, and omega-9 fatty acids were formulated into emulsions and were topically applied on rats with excision wounds. All omega-3-, omega-6-, and omega-9-rich oil formulations were found to accelerate wound closure compared to untreated, with significant improvement (p < 0.05) being observed at day 14. EPO induced early deposition of collagen as evaluated by Masson trichrome staining that correlated well with the hydroxyproline content assay, with the highest level at days 3 and 7. Vascular endothelial growth factor (VEGF) showed greater amount of new microvasculature formed in the EPO-treated group, while moderate improvement occurs in the LO and OO groups. EPO increased both the expression of proinflammatory cytokines and growth factors in the early stage of healing and declined at the later stage of healing. LO modulates the proinflammatory cytokines and chemokine but did not affect the growth factors. In contrast, OO induced the expression of growth factors rather than proinflammatory cytokines. These data suggest that LO, EPO, and OO emulsions promote wound healing but they accomplish this by different mechanisms.



The intent of this study was to test the effect of Top-D, a topical Vitamin D preparation, in delivering vitamin D.


Five hundred and fifty healthy patients, with vitamin D insufficiency and deficiency were recruited after written informed consent. Demographic data was recorded, adequate history and clinical examination was done to rule out any metabolic diseases. Complete blood picture, serum calcium, phosphorous, Parathormone and 25 Hydroxy-vitamin D3 (25OHD) was carried out before enrollment of the patients. Patients were divided randomly into two groups 350 in study group and 200 in the control group. Patients in the study group were given Top-D (Vitamin D3 gel made from proniosomal technology) to apply daily on the skin. Top-D 1 g contained 5000 IU of vitamin D3. The control group was given 1 g of Aloe vera gel to be applied every day. The two groups had no knowledge to which group they belong. After 4 months serum 25OHD was tested again.


Three hundred and forty five patients in study group and 192 in control group completed the study. The mean age of the patients in the both the groups was 42 years (18–80 years). The pretreatment 25OHD level in the study group was 11.03 ± 4.57 (2–12) ng/l compared to the control group 10.36 ± 4.09 (2–21) and post treatment the levels were 37.17 ± 6.04 (12–54) ng/ml and 10.51 ± 3.5 (2–19) ng/ml (p < 0.001).


The results of this study indicate that transdermal route of vitamin D is potentially, safe and can give desired results to raise the vitamin D levels. This route is an alternate route for supplementation of vitamin D which should be utilized


A new alginate dressing with Aloe vera gel was developed and used to study its cutaneous healing effect in vivo by surgical wounds in Wistar rats. The aloe-alginate film was characterized by Fourier-transform infrared spectroscopy, mechanical properties, scanning electron microscopy, hydration rate, and calcium release. The aloe-alginate film demonstrated desirable physical and mechanical characteristics for wound dressing application. The animals were divided in three groups (n = 20): gauze bandage, alginate film, and aloe-alginate film. The histological analysis on the 3rd, 7th, 14th, and 22nd days after the treatment showed that the aloe-alginate films modulated the inflammatory phase and decreased the quantity of macrophages when compared to the alginate film group and the control (gauze bandage). The evaluation of collagen fibers showed a decrease in type III and increase in type I fibers on group treated with aloe-alginate film suggesting an improvement in the progression of the healing process.

Therefore, topical administra- tion of vitamin D3 compounds may be an effective and safe route
to treat neuropathol- ogies of the skin. In conclusion, active vitamin D3 com- pounds may be a
potential therapy for periph- eral nervous disorders, stimulating NGF pro- duction in the

#2 Ruth

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Posted 10 July 2018 - 06:12 AM

Patent number 69696969: Aloe vera gel mixture of vitamin D3, DHA and doxycycline with added gelatin and ascorbic acid as only additives.

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