Newsletter of ARMARC Vol 28 Series 6 2 JUNE - 2022 ISSN: 2455-1384

A Comparative Wound Healing Study of Potoladi Ghrita, Jathyadi Ghrita and

Positive Control on Rats
* Dr. Suhas H.D., P.G. Scholar, Department of RSBK, ALNRMAMC, Koppa
Porf. (Dr). Dinesh Kumar Mishra, HOD, Dept. of RSBK, ALNRMAMC, Koppa
Dr. Jagadeesh Maiyya, Associate Professor, Dept. of RSBK, ALNRMAMC, Koppa
*Corresponding author: vdsuhas29@gmail.com
Received on: 21-04-2022 Accepted: 5-05-2022 Corrected: 25-05-2022
Abstract: Background: Wound is known to human beings since the evolution. Learning of wound healings
was result of observations of animals and nature. Disruption of integrity of skin, mucous, membranes and
organ tissues due to trauma, infection or any other reasons is referred to as wound. Healing of such wounds
is complex phenomenon involving physiological functions and medication at greater level.
Materials and Methods: 36 albino rats were randomly selected and they were divided in three groups named
as positive control, Jathyadi ghrita and Patoladi ghrita Methods selected for the study were excision wound
model and incision wound models. Assessment criteria was based on wound contraction and period of
epithelization. To check the role of collagen, tensile strength was measured.
Result: The mean and standard deviation for percentage of area of wound closure were respectively noted
on 4th, 8th, 12th and 16th days for Patoladi ghrita as 32.96 + 7.48, 54.40 + 14.16, 62.36 + 12.03and 77.55 +
6.33. The comparison between Patoladi ghrita and Jathyadi ghrita was not significant for 8th day and 12th
day measurement.
Discussion: Tensile strength and period epithelization were observed better in Patoladi ghrita group in
comparison to both Jathyadi ghrita and positive control. The component drugs of Patoladi ghrita are prominent
in Thiktha, Kashaya, Katu and Madhura rasa.
Conclusion: The Patoladi ghrita has shown higher healing rate and tensile strength in comparison to Jathyadi
ghrita and positive control. Positive control was having minimum in studied three groups.
Key words: Patoladi Ghritha, Jathyadi Ghritha, Vruna, Incision wound, Excision wound.
Wound is known to human beings since the dawn of humanity1 and almost all old civilizations have referred
to learning of wound healing with the observations to nature and animals. Later, responsibility came to healers.
Online medical dictionary defines wound as, “an injury or damage, usually restricted to those caused by physical
means with disruption of normal continuity of structures2. It is disruption of the integrity of skin, mucous, membranes
and organ tissues3. Charak Samhita Chikitsasthana 25/5 mentions two types of wound as innate and exogenous4.
These two types are further classified in twenty types on the basis of distinctive features5. Wounds of skin and
muscles are referred to easily curable wound, if it is recent in origin6. Curable means wound healing.
So, wound healing is a process of rectifying such disruption caused by trauma, infection, radiation etc. The
process of wound healing is a complex series of reactions and interactions with various conditions, so it differs as per
individual physiological bodies as many factors affect this including age, sex hormones, stress, diabetes, medication,
alcoholism etc.7 A normal wound healing is consisting of three successive phases hemostasis/inflammatory phase,
proliferative phase and remodeling phase5. Repair of damaged tissue occurs by regeneration and maturation of
uninjured cells, and deposition of connective tissue (primarily of collagen) to form a scar9.
Many formulations are suggested in Ayurveda for wound healing. These formulations are based on covering
all aspects including the initiation and activation of healing to infection control. Bases of number of these formulations
are ghrita. Some workers have worked on Jatyadi ghrita. So, present work was selected to compare the wound
healing properties of Patoladi ghrita with Jatyadi ghrita and positive control and three groups’ study.
Newsletter of ARMARC Vol 28 Series 6 3 JUNE - 2022 ISSN: 2455-1384
2. Materials and Methods
Reference for Preparation: Ashtangahridaya, Uttarsthana: 13/6-9
Procurement of Drugs: Leaves of Nimba, seeds of Ela, heartwood of khadira, fruits of haritaki and root bala were
procured locally from the forest. Remaining ingredients were procured from Udupi market.
Genuineness of Drugs: Genuineness of herbal drugs were confirmed by quality control laboratory of ALNRMAMC,
Koppa.
Manufacturing of Formulations: Pharmacy of ALNRMAMC, Koppa.
Place of Animal Study: Animal House, ALNRMAMC, Koppa.
Ethical Committee certification: ALNRAO/IAEC/Re/S/9/2020-21
Materials required for Animal Study: Diethyl ether as a vaporizing anesthesia, glass chamber, laboratory rat
cages, scalpel and blade, artery forceps, needle holder, non-absorbable sutures, surgical cotton, gauze, gloves,
povidone iodine, jatyadi ghrita, patoladi ghrita etc.
Care of animals: The rats in Animal house were purchased from In Vivo Biosciences, Bangaluru. The rats were
fed with Amruth brand rat pellets and purified water with reverse osmosis. They were kept in individual cages at 24-
26°C with light-dark cycle of 14-10 hours.
Inclusion criteria:
 Healthy albino rats of either sex
 Albino rats weighing 150-200 gm
Exclusion criteria:
 Infected and pregnant albino rats
 Rats used for other studies
 Rats below weighing below 150 gm and above 200 gm
 Rats showing signs of infection
Grouping of animals: 36 albino rats were randomly selected and they were divided in three groups. The groups
were named as positive control for povadine iodine ointment group, JG for Jatydai ghrita group and PG for
Patoladi ghrita group. Each group was further sub-divided in to two for excision and incision models.
Methods of animal experiment
10,11
Excision wound model : Initially rats were shifted to the glass chamber where cotton swab dipped in Di Ethyl
Ether was kept. After inhaling the anaesthesia for 5-10 minutes rats were anaesthetized. Anaesthetized rats were
kept in dissection plate, hairs on the dorsal surface of rats were removed using Veet hair removal cream and part was
cleaned with Betadine solution.
A round seal of 2.5cms in diameter was impressed on the Dorsal Thoracic Central region 5cms away from
the ear pinna of the rats. Full skin thickness from the marked area was excised in circular manner with the help of
forceps and surgical blade. Approximate area of 500mm2 was formed; once haemostasis was achieved rats were
shifted to individual cages.
Incision wound model11,12: Rats were anaesthetized same way as done excision method. Anesthetized rats were
secured to dissection plate in prone position. Hairs were removed using Veet hair removal cream from dorsal part
one-inch parallel to vertebral column. Area was cleaned with Betadine solution before incision. A paravertebral inc-
Newsletter of ARMARC Vol 28 Series 6 4 JUNE - 2022 ISSN: 2455-1384
-ision was made measuring 6 cm in length and of full thickness of skin. Interrupted sutures were put using non
absorbable sutures at an interval of 1 cm. Once haemostasis was achieved, rats were shifted to individual cages.
Post-operative stage: External application of povadine iodine ointmentwas applied on rats of positive control
group. Jathyadi Ghritha application was done externally to the rats of JG group while Patoladi Ghritha was
applied externally to rats of PG group. The wound was cleaned with normal saline and, povadine iodine, Jathyadi
Ghritha and Patoladi Ghritha were applied to rats of respective groups for 10 days. All rats were given normal food
and water. Cages were cleaned every day to avoid infection from their own faecal matters.
Assessment criteria: In excision wound model, wound contraction rate and epithelisation was assessed using
planimetry. In incision wound model, to study the role of collagen in wound healing, tensile strength of the wound is
assessed using densitometry.
Wound contraction: A wound heals by contraction, so to assess the contraction rate of the wound, wound margins
were measured using a transparent polythene sheets and subsequently retracing them on a millimetre graph paper.
This was later calculated as percentage of original wound size for each animal in group depending on days taken for
complete wound contraction.
Period of epithelisation: This was measured in terms of days required for falling of scar. Falling of scar leaving no
area behind was taken at the end of complete epithelisation and time noted in all albino rats.
Collagen strength: Here the tensile strength was measured in terms of breaking strength13. Volume of water
required to break the edges of the wound was measured and converted to corresponding weights assuming the
density equal to one. Using this tensiometer, the tensile strength is expressed as the minimum weight of water necessary
to bring gaping of the wound. In this method 8th day sutures were removed and tensile strength was determined on
10th post wounding day.
All the animals were anesthetized before measuring the breaking strength as tensile properties are usually
equated with tensile properties with collagen content. Each rats were secured to operation table in its supine position.
Two lines were drawn on the either sides of the wound, 3mm away from the wound margins on the adjacent normal
skin. Two allied forceps were hooked to a metal rod and it was fixed to a light polythene container through a string
run a pulley. Water was allowed to flow in constant rate into polythene container to build constant pulling through
forceps to disrupt the wound. The flow of water was regulated by means of an occlusion clamp on rubber tubing
connected to water reservoir kept in a suitable height. As soon as gaping of the wound was observed, water flow
was arrested to avoid further gaping of the wound. The wound was re-sutured and allowed to heal without sacrificing
the albino rats.
Statistical anlysis: ANOVA followed by Dunnett t-test
3. Result
Table Number: 1
Percentage of area of excision wound closure in mm square at different days
Group 4th Day 8th Day 12th Day 16th Day
M ean + St. dev. Mean + St. dev. Mean + St. dev. Mean + St. dev.
Positive control 32.96 + 7.48 54.40 + 14.16 62.36 + 12.03 77.55 + 6.33
Jathyadi ghrita 33.93 + 5.74 59.16 + 8.32 68.68 + 7.60 80.40 + 5.86
Patoladi ghrita 54.26 + 6.97 69.46 + 3.64 79.00 + 2.66 90.73 + 4.77
Newsletter of ARMARC Vol 28 Series 6 5 JUNE - 2022 ISSN: 2455-1384
Table Number: 2
Significant values between groups on different days for percentage area of wound closure
Groups 4th Day 8th Day 12th Day 16th Day
Positive control-Jathyadi ghrita > 0.05 > 0.05 > 0.05 > 0.05
Positive control-Patoladi ghrita < 0.05 > 0.05 < 0.05 < 0.05
Patiladi ghrita-Jathyadi gthrita < 0.05 > 0.05 > 0.05 < 0.05
Table Number: 3
Tensile strength (in weight) and Period of epithelization (in days)
Group Tensile strength (in weight) Period of epithelization (in days)
M ean + St. dev. Mean + St. dev.
Positive control 426.66 + 25.82 18.33 + 1.21
Jathyadi ghrita 431.66 + 31.25 18.00 + 0.89
Patoladi ghrita 480.00 + 38.06 16.33 + 1.24
Table Number: 4
Significant values between groups for tensile strength and period of epithelization
Groups Tensile strength Period of epithelization
Positive control-Jathyadi ghrita > 0.05 > 0.05
Positive control-Patoladi ghrita < 0.05 < 0.05
Patiladi ghrita-Jathyadi gthrita > 0.05 < 0.05
4. Discussion
The mean of the percentage of area of excision wound closure (mm2) was observed higher in Patoladi
ghrita in comparison to Jathyadi ghrita and positive control for 4th day, 8th day, 12th day and 16th being 54.26,
69.46, 79.00 and 90.73. Jathyadi ghrita group had shown comparatively higher side of mean of wound closure
than positive control. Even the tensile strength and period of epithelization also exhibited similar pattern as maximum
weight was tolerated in Patoladi ghrita group followed by Jathyadi ghrita group and positive control. The period
epithelization was minimum in Patoldai ghrita and maximum in positive control. Ayurveda has its own principle to
justify this differences as the rasapanchaka of the drugs used in Patoladi Ghritha gives us an idea of probable
mode of action.
Patoladi Ghritha has total 18 ingredients most of them have Thiktha, Kashaya, Katu and Madhura rasa
as prominent rasas. Thiktha and Kashaya rasa are mainly kledahara in nature and helps in Vrana shoshana14,15
and enhances the rate wound healing. Kashaya rasa having sroto vishodhana guna15 means clears the blocks in
the capillaries and improves circulation. Stambhana15 property of Kashaya rasa helps in achieving haemostasis.
Katu rasa helps in reducing the swelling caused by Vrana16. Madhura rasa helps in Sandhana means joining the
edges of the wound improves the tensile strength of wound edges17.
Most of the drugs have properties like Laghu and Ruksha, Laghu guna has a property of Kaphaghna and
does sroto shodhana means clears the blocks in the channels and aid in proliferation of surrounding connective
tissue elements. Ruksha guna helps in keeping the wound dry and helps in wound contraction.
Newsletter of ARMARC Vol 28 Series 6 6 JUNE - 2022 ISSN: 2455-1384
The drugs like Nimba, Patola, Musta, Chandana, Khadira has properties like Kanduhara, Vedana Sthapana
hence, helps in reducing the pain as well as maintaining the dryness in the wound. Vidanga, Nimba, Kutaja, Musta
and Khadira have anti-microbial and anti-bacterial properties and helps in preventing bacterial infections. Ghrita is
used as base for the formulation, however, it is having a property called Samskarasyanuvarthanam8 means, it
takes the properties of dravyas used during Samskara at the same time doesn’t lose its original properties. Ghritha
is having the propery of Vrana Ropana. It also acts as rasayana, so along with qualities of ghritha it also acts as
Kledahara, Kanduhara, Vranaghna, Shothahara as these properties are inherited by dravyas used in the
formulation.
5. Conclusion
The Patoladi ghrita has shown higher healing rate in terms of percentage of area of excision wound closure
(mm2) on 4th day, 8th day, 12th day and 16th compared to Jathyadi ghrita and positive control. Even the tensile
strength was better in Patoladi ghrita in comparison to Jathyadi ghrita and positive control. Positive control was
having minimum in studied three groups. Period of epithelization was maximum in positive control and minimum in
Patoladi ghrita group. Further detailed studies are required based histopathological screenings.
Conflict of Interest: No conflict of interest lies as per authors.
Funding: Not funded
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6. Ibid. Sharma, P.V. (1998). Charak Samhita. Chikitsasthana: 25/36-37. p.411.
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Chinese Medical Association : JCMA, 81(2), 94–101. https://doi.org/10.1016/j.jcma.2017.11.002
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internationales de pharmacodynamie et de therapie, 196(1), 117–126.
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Resina Draconis (Dracaena cochinchinensis) in Animal Models. Evidence-based complementary and alternative
medicine : eCAM, 2013, 709865. https://doi.org/10.1155/2013/709865.
12. Ehrlich, H. P., & Hunt, T. K. (1969). The effects of cortisone and anabolic steroids on the tensile strength of
healing wounds. Annals of surgery, 170(2), 203–206. https://doi.org/10.1097/00000658-196908000-00007
13. Rovee D.T., & Miller C.A. (1968). Epidermal Role in the Breaking Strength of Wounds. ArchSurg. 1968;96(1):43–
52. doi:10.1001/archsurg.1968.01330190045011
14. Vidyanath, R. (2013). Trans. Illustrated Ashtanga Hridaya of Vagabhata. Sutrasthasthana. Sutrasthana 10/
15. p.176. Chaukhamba Surabharti Prakshan, Varanasi.
15. Ibid. Vidyanath, R. (2013). Sutrasthana 10/20-21. p.177. Chaukhamba Surabharti Prakshan, Varanasi.
16. Ibid. Vidyanath, R. (2013). Sutrasthana 10/17. p.176. Chaukhamba Surabharti Prakshan, Varanasi.
17. Ibid. Vidyanath, R. (2013). Sutrasthana 10/7-9. pp.174-175. Chaukhamba Surabharti Prakshan, Varanasi.
18. Ibid. Vidyanath, R. (2013). Sutrasthana 16/2. p.255. Chaukhamba Surabharti Prakshan, Varanasi.
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