Sharma, A.K.Kumar Singh, Saurav2019-01-162019-01-162018http://krishikosh.egranth.ac.in/handle/1/5810090755EVALUATION OF WOUND HEALING POTENTIAL OF PLATELETS RICH PLASMA WITH OR WITHOUT ULTRAVIOLET THERAPY IN DOGSThe present experiment was conducted on 90 surgically created wound in fifteen apparently healthy dogs of either sex of one and two year .The selected dogs were dewormed two week prior to commencement of experiment. All dogs were maintained under iso-managemental condition and similar feeding schedule. The wounds of groups I,II and III were treated with PRP, PRP+PRPgel and PRP+Ultraviolet ray respectively. Gross morphological parameter such as general appearance of wound, appearance of granulation tissue, degree of inflammation ,filling of wound gap by granulation tissue, extent of percentage wound contraction, appearance of scab and shedding, complete healing and extent of cicatrisation were studied on day 3, 9, 15, and 21 post treatment. Physiological parameters like heart rate (beats/min), respiration rate (breaths/min) and rectal temperature (oF) were recorded before and at 3, 9,15, and 21 days post treatment. Histomorphological parameter like inflammatory cell infiltration, neovascularization, fibroplasia, collagen fibres and epithelialization were studied on day 3,9,15 and 21day. Histochemical parameter for wound healing viz. onset and extent of collagen fibres, elastin fibres and reticular fibres were observed at interval of 3, 9,15 and 21 days post treatment. These parameters were visually quantified on the scale of 0,1,2,3 and in some parameter up to 4. There was Clotting of blood on day 1st post wounding in all the groups. On 2rd days the wound appear moist and pinkish in all the groups. On 4th days the wounds were exudating in PRP and PRP Gel treated group whereas no exudation has been found in the UV treated group. Scabbing was started in 5th to 6th days interval of post wounding in all the groups which were completed between 11st to 14th days in UV and PRP group as compared to PRP Gel .Sloughing completed on 19th to 21stdays in the wounds treated with PRP alone (group I) and in combination with UV rays (Group III). Whereas, PRP gel group (group II) exhibited moderate sloughing by the end of observation (21 days). In PRP group, wound contraction was progressive and significantly higher at different interval of as compared to wound contraction on 3rd day. Almost similar observation has been found in other two groups also but the maximum contraction of significance (P<0.05) were recorded on 9th and 15th days post treatment in ultraviolet group as compared to PRP Gel and PRP solution alone. A Transient and non- significant (p>0.05)variations in heart rate has been recorded at different intervals in all the groups .Respiration rate recorded at different interval of observation before and after treatment in all the groups showed non significant variations (p>0.05). Rectal temperature on 3rd day post wounding were (101.74±0.24), (101.04±0.26) and (101.4±0.15) oF in PRP solution, PRP Gel and ultraviolet groups, respectively (p>0.05). From 3rd onwards, the value of rectal temperature did not differ significantly within and among the groups. Leucocytic infiltration, neovascularization, fibroplacia, epithelialization, collagen formation, elastin formation and reticular formation has been observed histomorphologically at different intervals of observation post treatment with PRP solution alone or in combination of PRP gel or Ultraviolet therapy in the experimental wounds of dogs. Leucocytic infiltration showed marked and significant at 3 days of observation in all the groups. Neovascularization and epithelialization exhibited marked enhancement in all the groups at 9 and 15th days of observations, whereas in other histomorphological parameters marked with high and significant scoring mostly at 15 and 21st days of observation post treatment. As compared to PRP solution and PRP gel groups, UV group exhibited more marked alterations in favour of wound healing on 3rd day of histophathological observation, however the extent of regenerative epithelialization was clearly observed to be maximum in the wound treated with PRP +UV followed by PRP +PRP GEL and lowest in those treated with PRP alone . Granulation tissue at this period of observation were composed of plump fibroblast cells scattered within fine collagen fibres ,thin walled new capillaries and mononuclear cells admixed with oedematus and inflammatory changes in all the groups with no microscopic differences of significance among them. On 9th day of observation, epithelialization and granulation became more marked along with intense appearance of fibroblast, collagen and capillaries in UV treated wound followed by PRP solution and PRP gel. There was continued increase in accumulation of collagen fibres and proliferation of fibroblast from 15 to 21 days. Histochemically, microscopic section in group I and II showed the moderate appearance of collagen and elastin and reticulin fibers. However increase in the amount of the matured collagen, elastin and reticulin fibres were more pronounced in wound treated with PRP+UV followed by PRP+PRP GEL and PRP alone, respectively with reduction in leucocytic infiltration and neovascularization. Among the group , ultraviolet treated wound exhibited significantly higher (p<0.05) SOD level in homogenised tissue from 3rd day onward as compared to other groups at corresponding interval. However the catalase value observed at 21st days showed significant fall (p<0.05) as compared to the value recorded at different interval in all the groups. The maximum elevation of SOD and catalase were recorded at 15th days of observation in ultraviolet group as compared to other groups. The values of catalase activity recorded at different interval showed a non-significant variation among the groups. The MDA activities exhibited progressive and significant decrease (p<0.05) at different period of observation in all the groups compared to the base value. The value of MDA activity in ultraviolet group showed maximum decrease as compared to other groups with corresponding intervals. On the basis of above findings following conclusions can be drawn: • Preparation of platelet rich plasma (PRP) solution and PRP gel was quite easy, economical and with need of limited resources. • PRP solution alone and in combination with PRP gel or ultraviolet therapy had significant effect in reducing inflammation and increase in granulation of wound. These were more pronounced in ultraviolet treated group followed by PRP solution and PRP gel. • Wound treated with PRP solution along with ultraviolet exhibited highly significant wound contraction at 9 and 15th days of observation as compared to PRP solution alone and PRP solution with PRP gel. • Heart rate, respiration rate and rectal temperature were non- significantly variables at different intervals of observation in all the groups. • Fibroplasia, epithelialization and neovascularization appeared more in ultraviolet group followed by PRP gel and PRP solution. • Wound treated with PRP solution in combination with ultraviolet proved to be superior in terms of early increase in collagen, elastin and reticulin fibers. • Super oxide dismutase (SOD) and catalase activity in all the groups showed a progressive and time dependent increase. However, SOD activity was more marked and significant in ultraviolet treated groups. In contrast of SOD and catalase, the MDA (Melondialdehyde) activity was progressively decreased at different intervals of observation in all the groups. A marked reduction in MDA activity was recorded in PRP gel at 15 days and in ultraviolet group at 21 days post wounding. • During the present study the intradermal administration of autologous PRP solution at wound margin and at the center of wound along with topical application of PRP gel or ultraviolet therapy showed significant enhancement of wound epithelialization and reduced scar formation. PRP solution in combination of ultraviolet therapy showed marked decrease in oxidative stress, extensive wound contraction, diminution of scar and early maturation evidenced by extensive collagen, reticulin and elastin fibers. • Hence, intradermal injection of PRP solution followed by ultraviolet therapy can be recommended as better enhancement of wound healing in dogs over PRP solution alone or in combination of PRP gel.en-USnullThesis