Frost bite produced in rates by exposing them to -15 degree calcius and the extent of injury in the hind limbs and the tail was assessed at the end of 15 days. Hyperoxic treatment at normal atmospheric pressure and 1.5 atmosphere was given to experimental groups for 30 minutes daily for seven days in a hyperoxic chamber immediately after cold exposure. The controls were not given any treatment. Single oxygen treatment at normal atmosphere pressure was of no value, however, repeated hyperoxic treatment showed limited improvement but repeated treatment for seven days with hyperbaric oxygen at 1.5 atmosphere showed distinct recovery of frozen parts

Hegde, K. S.

Krishna, Bal

Mathew, T. Lazar

English

Defence Science Journal

USE OF HYPERBARIC OXYGEN IN EXPERIMENTAL FROST-BITE BAL KRISHNA, K.S. HEGDE & LAZAR MATEEW Defence Institute of Physiology & Allied Sciences, Delhi Cantt. (Received on 21 January 1977; revised 27 June 1977) Brost-bite was produced in rats by exposing them to -15OC and the extent of injury in the hindlimbs and the tail was assessed at  the end of 15 days. Hyperoxic treatment at  normal atmospherio pressure and 1'5 atmosphere was given to experimental groups for 30 minutes daily for seven days in a hyperoxio chamber immediately after cold exposure. The controls were not given any treatment. Single oxygen treatment at  normal atmospheric pressure was of no value, however, repeated hyperoxic treatment showed limited improvement but repeated treatment for seven days with hyperbaric oxygen at  1 .5 atmosphere showed distinct recovery of frozen parts. Extensive studies have been carried aut far the prevention and treatment of cold injuries, but the prablem still remains unsolved. Hyperbaric axygen (OHP) treatment far cald injuries may be considered optimistic. While a number of experimental studies reparted the effectiveness of OHP therapy in healing and tissue survivall~2, this was, hawever, nat corroborated by subsequent observations. The present study was undertaken to evaluate the effect of OHP therapy as a treatment for cold injury. E X P E R I M E N T A L  . Cald injury was praduced in five graups k~f male rats, each group consisting of 15 rats. -The body weight of the rats ranged from 172 to 240 gm. with a mean of 208 -J= 1.89 gm. The rats were expased to - 15°C f 1°C in a thermastatically cantrolled deep freeze. The groups were made by random selection. -The experi- ments were conducted in two series. Series A : Three rats, ane each from graup 1, 2 & 3 were exposed to cald simultaneously, restrained in individual pelspex harness with the tail protected4, At the end of the cold exposure the rats were taken . out, the rat fram graup ane was kept at raom temperature and served as cantrol. The ather two were - treated in the hyperoxic chamber for 30 minutes at atmospheric pressure. The rat fram graup two, received treatment only for one day and that from group three, received treatment for 30 minutes daily for seven days. Fifteen such experiments were canducted. 8eries B : Two rats, one each from graup 4 and b were simultaneously exposed to -15°C & 1°C in a metallic cantainer for three hours. Air circulation was maintained through a rubber tube and a circu- lation pump kept outside the deep freeze. The rats were free to move and the tail was not protected in order to assess the degree of injury in the tail as well. During expo;iure the rat assumes a posture to reduce its surface area and also the heat loss fram the body. At the end af the experiment the rat from group four was kept at room temperature and served as control and the other from group five was given hyperbaric oxygen treatment at 1.5 atmospheric pressure for 30 minutes daily, far seven days. Pifteen such experiments were conducted. The thawing in all animals was done at  room temperature (20-25OC). After the treatment the rats were kept in experimental room for a period of one haw and then the legs were bandaged using dry sterile ' cotton to prevent infection and mechanical damage. The rats were transferred to the animal house which is DEF. SIX. J., VOL. 28, APRE 1978 maintained at  20-25OC. The bandages were replaced daily for a period of 15 days at the end of which the degree of injury was assessed using the following criteria6. Very sevem, loss of enthe paw or more (++++); Severe, loss of all the digits (+++); Mode- rate, loss of two phalanges or equivalent (++); Primary, minimal injury without loss, of pha- langes (+) and No tissue damage (0). The hyperoxic and hyperbaric environment in the exposure chamber (volume 16.5 litres) was obtained by introducing oxygen from a pressurised axygea cylinder. No water vapour was added to the system. R E S U L T S  A N D  DISCUSSION The data of the A series of experiments (hyperoxic treatment) is given in Table 1. Statistical analysis has been done by Manu-Whitney U Test, noting the degree of injury expressed in numbers for each rat in all the groups. f3tatistical comparison in controls of series A and B shows no s i m c a n t  difference. It is seen that there is no signi6caut difference in recovery between gr~ups  1 and 2 but group 3 (hyperoxic) treated for seven days shows significant (P < 0.02) improvement compared to control group 1. TABLE 1 %XTBCT OF HYPEROXIU TREATMENT IN EXPERIMENTAL F&oST-BITE 81. Group 1 (Control) Group 2 Group 3 No. Without 0, treatment O, treatment-only for one day 0, treatment for seven days r A r --Lt Paw Rt Paw Lt Paw Rt Peff Lt Paw Rt Paw , The statietbal oompariaon showa no eignificant digerenee between group 1 and 2. A signScant difference (P < 02) is between group 1 a d  3. BAL KRISHNA, et d. : Hyperbario Oxygen in Frost-Bite In the B series of experiment in which hyperbaric treatment was given, there was highly significant difference (P < 0.001) in recovery between the experiments1 ar~d control groups (Table 2). Group 4 (Control) Group 5 (Experimental group) SI. Without 0, treatment 0, treatment at 1 .5  atm. for s e v e ~  days NO. c -----> r .  .A > Lt Paw Rt Paw Tail affeoted (%) Lt Paw Rt Paw Tail affected (%) 14 + + 30.0 0 0 0 1.4 -F+ f + 47.7 f + 30'2 Mean 66'4% Mean 8'8% Stati~tical oompariaon shows signifimnt (P < 0'001) difference between the  group^. The main oonolnsions derived from the data are (a) Hyperoxic treatment for a single day at atmospheric pre8sul:e does not enhance the recovery of the frozen parts in the experimental rats but if the treatment is repeated for seven days, a marginal improvement is seen. (ii) Hyperbario oxygen therapy at 1.6 atmos- phere leads to siyi6cant recovery of the frozen parts when the treatment is continued for seven dap. Frost-bite, the severest type of cold injury results from actual freezing of the t h e s .  Freezing below 4 to - 7 O C  crystallises cell protoplasm which results in tis$ue damage. During thawing, blocking of blood vesselel' obstructs the circulatian resulting in conglutinatioa6. Liberation of histamine like substances, causes increased permeability of the capillary wall, collection of exudates in tissue spaces, edema and blisters'. In the find stage degeneration of red blood cells leads to hyaline mass production and the tissue damage is due to hypoxia. DEF. Bur. J., VOL. 28, ~ R I L  1978 Necrosis is primarily the result of vascular stasis and anoxemia and is not influenced by colds. It is reported that hyperbaric treatment improves tissue oxygen contentg. With its bacteriastatic action hyper- baric oxygenation may accelerate the tissue regeneration 9, 10. In  our study it is believed that hyperbaric oxygen treatment may increase the amount of oxygen dissolved in the blood, thereby supplementing more of oxygen which relieves tissue hypoxia. A similar effect may be achieved ta a lesser extent, by repeated hyperoxic treatment as seen in group 3. Further, the hyperbaric axygen treatment may lead to  epi- thellal regeneration, and capillary proliferatianxl. When oxygen is administered under pressure, i t  increases the oxygen saturation of the blood plasma which helps the affected cells to receive the requisite amaunt of oxygen during thawing. In the present study, OHP treatment was given immediately after freezing of the paw. Okuboj e et al.1, used similar procedme for hyperbaric oxygen treatment in rabbits. Our results of OHP therapy in series B support their finding. On the other hand Hardenberg working on mice with the paw exposed to -30°C for 1-2 min., treated them with OHP, 3-4 hours after freezings. The cold expasure was rather of a very severe degree and the damage to the tissues might have been complete and could not be reversed by OHP. The recovery by hyperbaric oxygen in our experiments shows beneficial effect of OHP in the prevention of ccjld injury of moderate to severe intensity when the therapy is instituted early. It eliminates the aggra- vating influence of hypaxaemia on tissue damage. Signdieant degree of recovery is possible when the the- rapy is instituted early and when the tissue damage is n ~ t  complete. R E F E R E N C E S  1. OEUBOYE, J.A. & FERUUSON, C.C., Can. J. Surg., I1 (1968), 78. - 2. SMITE, G., Dia. Cheat, 45 (1964), 15. 3. HARDENBERUE, .J., Surg. Rea., 12 j1972), 34. 4. B ~ L  ~ S H N A ,  CKATURVEDI, R.C. & BHATIA, B., Prediction of snsceptibility to frost-bits, part 111-'Experimental Frost-Bite in Rate'. DIPAS Report No. 2/66, AFMRC Project No. 152164. 6. LAZm MATHEW, PURKAYASTHA, S.S. & RAI, R.M., Indian J. Ezp. Biol., 11 (1973), 230. 6. KREYBERU, L., Lancet, 1 (1946), 338. 7. LEWIS, T., Brit. Med. J., 2 (1941), 796. 8. K~EYBERG, L.,TOM, F., WHAYNE, T.E., MICHAEL & DEBAKEY, E., 'Cold Injury Ground Type' (Department of Army, Washington, D.C.), 1958, p. 239. 9. IKEDA, K., h m 1 ,  H., NAGAO, H., KAI~INo, K., SUGLI, S., IWA, T. & WADA, J., 'Proc. IV Intl. Cong. Hyperbaric Medicine', (Igaku Shoin Ltd., Tokyo), 1969, p. 377. 10. KETOHUM, S.A., THOMAS, A.N. & W, A.D., 'PPOC. IV Intl. Cong. Hyperbaric Medicine', (Igaku Shoin Ltd., Tokyo), 1969, p. 388. 11. Z I M ~ B ,  J.G. & DEXNIS, D.J., Xcienee, 140 (1963), 994. 

Use of Hyperbaric Oxygen in Experimental Frost-Bite

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Use of Hyperbaric Oxygen in Experimental Frost-Bite

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