Seabuckthorn and Dogs Study

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Seabuckthorn, a resource for soft tissue repair in animals

A.C. Varshney1and S.P. Tyagi2

1Professor and Head, 2Assistant Professor

Department of Surgery and Radiology, College of Veterinary and Animal Sciences

CSKHPKV, Palampur, H.P., India

vetsurg@hillagric.ernet.i

            Wound healing is a complex phenomenon involving a number of finely synchronized cellular processes of body which are in turn governed by a number of control systems of the body. However, a number of variables affect this normal response of body in many ways. In general, the availability of various vitamins, minerals, fatty acids, amino acids and many other nutrients in body have a major influence on this process directly and indirectly. Moreover, various disease processes and myriad external conditions also affect this process in an unanticipated manner. Therefore, management of any kind of wound is basically aimed in controlling the above mentioned variables in such a manner that the body functions optimally in its healing endeavor. The management of wounds in animals is always potentially complicated because of little control on these variables compared to human. Because of this, the wounds frequently turn chronic and ulcerative in animals, where this natural healing process is further disrupted. The scientists all over the world continuously strive to find better wound-healing-supporting medicines and means to shorten the wound healing period as much as possible. Moreover, in cases of chronic and ulcerative wounds, exogenous application of certain growth promoting agents or compound that can enhance in-situ generation of various growth factors becomes essential to augment the healing process. Taking cue from traditional medical practice of various places of world, scientists have tested many herbal preparations for this purpose by employing modern scientific methods. Due to the presence of a number of bioactive substances sometimes possibly in a synergistic ratio useful for wound healing process, many such herbal preparations have shown promising results. Already a number of herbal drug companies are marketing various wound healing medicaments for animals and human the world over.  Seabuckthorn (Hippophae sp.) is one such useful plant which is already known for its various therapeutic applications because of presence of many bioactive compounds mainly in its fruit and seed. The fruits of seabuckthorn (Hippophae rhamnoides L) have been used as a drug by traditional Tibetan and Mongolian medicine since ancient times. It had pharmacological effects on the lungs, stomach, spleen and blood circulation, which were recorded in some medicinal classics, such as Sibu Yidian from the Tang Dynasty and Jing Zhu Ben Cao from the Qing Dynasty. Seabuckthorn has also been mentioned in the writings of ancient Greek scholars such as Dioscorid and Therophast. In modern times, however the plant attracted the interest of scientists mainly only after the 1977 when the Ministry of Public Health, China officially listed seabuckthorn in the Chinese Pharmacopoeia for the first time (Singh 2001).

China and Russia are forerunner in exploiting the potential of seabuckthorn as medicinal plant. There this plant is being cultivated as medicinal cash crop in thousands of hectares of land. Russian scientists could not only be able to identify a number of key bioactive substances present in seabuckthorn but also substantiated their pharmacological actions through systematic research on experimental animals and also through many clinical trials on human beings as well (Mironov et al.,1989). Since 1985, scientists of many counties remained engaged in a series of scientific experiments on its juice, oil and other extracts with analyses of its nutrient and chemical composition, pharmacodynamics and toxicology. Taking leads from these developments, now over 40 countries of world are actively engaged in conducting research on this plant and harvesting its potential. A number of steroid, flavonoids and vitamins present in the plant are thought to be responsible for its versatile pharmacological activities such as anti-inflammatory (Zhang Wenlu et al., 1988), chemical or physical burn wound healing ability (Nikulin et al., 1992, Vlasov, 1970), anti-gastroulcerative activity (Xiao et al., 1992), hepatoprotective (Cheng, 1990, 1992), anti-cancerous (Li and Liu, 1991), anti-atherosclerotic (Olziikhutag, 1969), anti-lipemic and anti-arrhythmic (Fengming et al., 1989) properties. It has been reported that seabuckthorn contains more than 190 bioactive compounds in the seeds, pulp and juice. These compounds include many fat soluble vitamins (A, K and E), water soluble vitamins (C, B1, B2, folic acid etc.), 22 fatty acids, 42 lipids, organic acids, amino acids, carbohydrates, tocopherols, flavonoids, phenols, terpenes and tannins (Zhang,1990). The versatile pharmacological activities of seabuckthorn oil are hence attributed to these bioactive substances. The chromatographic analysis of ripe fruit of seabuckthorn revealed the presence of malic acid, oxalic acid and other unidentified acids in it. These organic acids are known for certain physiological functions in body such as reducing the toxic effects of some medicines like barbitals and antibiotics, preventing teratogenesis, damages from the X- rays and side effects of oxygen therapy. There is a distinct possibility that the available bioactive substances in seabuckthorn may help in reducing the inflammation, augment the tissue regeneration, epithelization and improvement in the microcirculation of blood capillaries in case of wounds thus hastening the overall healing (Qibikeva, 1989). Vitamin A helps in collagen synthesis and cross-linking of collagen fibres. In addition to this it also helps in proper epithelization. Vitamin E has anti-oxidant properties. Seabuckthorn also has flavonoids due to which it has anti-inflammatory properties and helps in promoting healing of wound and ulcer (Chen et al., 1988). Zinc helps in synthesis of proteins, RNA, DNA, fibroplasia, epithelization and cell mitosis. Magnesium decreases phagocytosis. The triterpene components of seabuckthorn oil (Ursolic Acid) has also been suggested to be responsible for wound healing, ulcer healing and in easing inflammation and pain (Ge, Xiaoyan et al., 1986). The exact mechanism responsible for augmenting wound healing by seabuckthorn flavonoids present in its oil remain speculative. Free radicals are generated at the site of injury, which impairs the healing process. Since seabuckthorn have anti-oxidative effects (Geetha et al, 2001) it is possible that healing activity of seabuckthorn might have been due to its potential to combat oxidative stress (Gupta et al., 2001). The different preparations of seabuckthorn such as decoction, powder, pill, medicinal extract, shortbread, ash and tincture have been used for the treatment of various disease conditions (Mingyu et al., 2001) but the main research emphasis remained focused on its oil.

Therefore, seabuckthorn seed oil has been used in a number of wound healing studies (Vlasov, 1970; Mironov et al., 1989; Buhatel et al., 1991; Khirurgia, 1995; Gupta et al., 2001, Mahajan, et. al. 2002, Varshney, et. al 2003, Kumar, et. al. 2003) in animals as well as in human. It has also been subjected to a number of anti-inflammatory studies in experimental animals and human (Lebedeva et al. 1989, Wu et al. 1992, Che et al. 1992, Varshney and Tyagi, 2004). Lebedeva et al. (1989) evaluated the efficacy of seabuckthorn oil in induced inflammation of mouse subcutaneous tissue and found its anti-inflammatory effects. Wu et al. (1992) studied seabuckthorn oil and oil embolus with seabuckthorn compounds for the treatment of chronic cervicitis and reported 97 percent general curative effect. Che et al. (1992) found therapeutic effects of seabuckthorn oil embolus on easing pain and eliminating inflammation. Li et al. (1989) reported anti-radiation effects of seabuckthorn oil. Fayman (1991) treated postoperative wound of tonsillitis with seabuckthorn oil. Xu Mingyu et al. (1993) described the role of seabuckthorn oil in eliminating inflammation and slough, easing pain, promoting immune function and strengthen body résistance. In a series of studies to evaluate the anti-inflammatory effects of seabuckthorn oil on experimentally induced acute and sub-acute cutaneous inflammation in dogs, Varshney and Tyagi (2004) found that seabuckthorn oil has mild anti-inflammatory effect on acute cutaneous inflammation. Tazakistan state hospital has studied the major biochemical mechanism of SBT oil’s anti-inflammatory activity. A triterpenoid compound, citroostadienol was identified in SBT oil, which had a strong anti-inflammatory activity. Platelet activating factor (PAF) plays an important role in acute inflammation and terpenoids and flavonoids found in SBT oil have marked inhibitory effect on PAF. Seabuckthorn oil was found to stimulate regeneration of tissues in rabbits to help achieving earlier wound healing (Mironov et al., 1989). Similarly the seabuckthorn oil was found to exert favourable effect on the process of cicatrisation during wound-healing in rats thereby reducing the total wound-healing period in another study by Buhatel et al.(1991). Seabuckthorn oil have also been shown to produce beneficial effects in the treatment of superficial burns of the skin (Vlasov, 1970) and operative wounds of ear, nose and throat (Fayman, 1991). Seabuckthorn oil was again found to be very effective by Nikulin et al., (1992) in the treatment of 1st, 2nd or 3rd degree chemical or physical burns as it helped in improving the formation of epithelia and reducing the healing period. Fuheng (1991) and Alieva (1976) also reported the use of seabuckthorn oil for the treatment of burns, skin radiation, cervical erosion, gastric and duodenal ulcers. Fan Yulin et al. (1991) used seabuckthorn oil in clinical treatment of traumatic perforation of tympanic membrane and reported full layer hyperplastic reunion of tympanic membrane.

Most of the research on seabuckthorn was however either done on laboratory animals such as guinea-pigs, rats and rabbits or directly on human patients. As far as domestic animals especially large and small domestic animals are concerned not much work has been done. Therefore, we have undertaken a research project to verify the wound healing-supporting abilities of seabuckthorn and its evaluation as dressing agent for cutaneous wounds in dogs, infected cutaneous wounds in calves in form of ointment and as oil in burn wounds in calves. More work is going on to evaluate the therapeutic efficacy of seabuckthorn oil on aseptic incisional and septic excisional wounds in calves and aseptic excisional wound in dogs. In the study to evaluate the efficacy of seabuckthorn ointment on the healing of aseptic wounds in dogs by Gupta et. al. (2002), the seabuckthorn ointment was compared with liquid paraffin and a commonly used antiseptic wound ointment 5% povidone-iodine. Grossly the signs of acute inflammatory reactions in cutaneous wounds were less pronounced and disappeared earlier both in seabuckthorn ointment and 5% povidone-iodine ointment treated animals. These signs subsided as early as on 3rd day as compared to around 7th day in paraffin treated wounds. Similarly the granulation tissue and scab formation in the wounds was noticed earlier in seabuckthorn and 5% povidone-iodine treated animals leading to early shedding of scab and achieving complete clinical healing within 14th –18th day as compared to control group of liquid paraffin treated wounds where it took about 21-24 days. The seabuckthorn and 5% povidone-iodine ointment treated wounds also showed early and sustained greater wound contraction of 36% and 32% respectively on 7th day and 75% and 77% respectively on 14th day as compared to 17% and 69 % at 7th and 14th day respectively in liquid paraffin treated wounds. Grossly the wound healing process progressed almost comparably in seabuckthorn and 5% povidone-iodine ointment treated wounds.  The histological observations of the healing tissue biopsies however, affirmed wound healing response in seabuckthorn ointment treated wounds even better than those treated by 5% povidone-iodine. It was evidenced by relative milder response of inflammation and greater fibroblastic proliferation in the wounds in the early stages of healing in seabuckthorn treated wounds. In later stages as well the process of epithelialization and the maturation and contraction of collagen fibers was found to be of most advanced nature in seabuckthorn treated animals.  The collagen fibres were arranged parallel to the skin surface in cutaneous wounds in this group as compared to other two groups where the mature fibrous tissue appeared loosely arranged and comparatively disoriented even at 28th day of healing.

In another study conducted by Mahajan et. al. (2002) to evaluate the efficacy of seabuckthorn in the healing of infected cutaneous wounds in calves, the gross wound healing response was found to be comparable in seabuckthorn and 5% povidone-iodine ointment treated wounds. Whereas, histological observations of the healing tissue biopsies revealed better wound healing response in the initial stages in 5% povidone-iodine treated wounds. It was evidenced by relatively greater angiogenesis, fibroblastic proliferation and lesser fibrinopurulent exudation in 5% povidone-iodine treated wounds. However in later stages the wound healing response was comparable in seabuckthorn and 5% povidone-iodine ointment treated wounds which were nevertheless better than liquid paraffin treated wounds.

Similarly the seabuckthorn seed oil was compared with 5% povidone-iodine ointment and liquid paraffin in the healing of burn wounds in calves by Kumar, et. al. (2003) and Varshney, et. al. (2003). It was again observed that the signs of acute inflammatory reactions in wounds remained less pronounced in seabuckthorn oil and 5% povidone iodine treated animals as compared to paraffin treated animals. The healing progressed fastest in seabuckthorn oil treated wounds as evidenced by earliest shedding of burn eschar in this group. Histopathological examination of the healing tissue biopsies also substantiated the faster progression of healing process in the seabuckthorn oil treated wounds. It was evidenced by presence of lesser neutrophils and mononuclear cells in the wound area, earlier and greater fibroblastic proliferation and its better organization towards the end of study period in seabuckthorn oil treated wounds.

The therapeutic and prophylactic efficacies of seabuckthorn oil on experimentally created gastric erosion/ulcers have also been evaluated in dogs at this institution. The group of animals that were fed seabuckthorn oil prophylactically showed lesser intensity of gastric erosions endoscopically in response to ulcerogenic drugs as compared to other animals. The gastric lesions also disappeared early in this as compared to other two groups. More research in this aspect is in progress.

It is therefore, concluded that seabuckthorn has a therapeutic potential for the management of various kinds of aseptic and infected cutaneous wounds, burns and gastric ulcers in animals.

References

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