Perception of postgraduate students in pharmacology toward animal simulation model.

OBJECTIVE: The objective of the study is to evaluate the perception of postgraduate pharmacology students toward computer-simulated method (CSM) in comparison to the prevalent isolated live tissue-based bioassay method.
MATERIALS AND METHODS: A questionnaire-based survey was conducted in 30 postgraduate pharmacology students who had used the animal simulation software and had completed at least five isolated tissue experiments. Students' opinions on the usage, logistics, advantages, disadvantages, and usefulness of CSM compared to live animal experiments (LAE) were analyzed.
RESULTS: Four tissues were used for LAE, whereas with CSM, students could perform experiments using 11 different tissues. Of the total nine bioassay methods, students had performed six assay methods using both LAE and CSM. Majority of the students (23/30) agreed that CSM reduces anxiety, technical errors and is less time consuming when used before LAE. Most of the students agreed that CSM can be used for difficult, lengthy experiments (19/30), and for UG/PG teaching (19/30). However, opinions regarding replacing LAE with CSM in PG teaching were divided (agree: 7, neutral: 12, and disagree: 12).
CONCLUSION: CSM should be integrated alongside LAE to complement, reinforce, and enhance learning from other techniques. Copyright:

Introduction

Pharmacology is the science of drugs which involves understanding the interaction of exogenously administered chemicals with living systems, including interactions between the drug molecules and the receptors. Animal experiments form an integral part of both, undergraduate and postgraduate pharmacology teaching. Animal experiments which include the demonstration of drug effects on isolated tissues or whole animal are an essential component of postgraduate practical pharmacology. However, these experiments are tedious, expensive, and time-consuming. Furthermore, the use of animals for teaching and learning of pharmacology has shown a downward trend over the last decade due to ethical concerns as well as because of practical problems associated with the animal experiments such as availability of animals, cost of purchasing animals, and maintaining animal houses.[1] The University Grant Commission (UGC) has decided to stop the animal experimentation for undergraduate students, citing serious ethical concerns related to the use of animals for experiments. Pressure from animal right organizations and subsequent guidelines by committee for the purpose of control and supervision of experiments on animals (CPCSEA) has seriously limited animal use in postgraduate training in experimental pharmacology.[2]

Replacement of animal experiments by computer-simulated model (CSM) has made pharmacology learning highly interactive and motivating. CSM improves students' critical thinking and clinical reasoning.[3] Various computer animal experiment simulated softwares are now being used for undergraduate and postgraduate practical teaching in various medical institutes of India. However, many academicians debate that learning experimental pharmacology by taking live tissues, if possible, will be more beneficial than learning on simulation models, which does not give a hands-on experience.

Although many studies have explored the perceptions of the CSM in undergraduate medical education, the literature for the postgraduate students' perceptions regarding the use of CSM in postgraduate training for isolated tissue experiments have not been investigated.[456] Thus, it was deemed essential to assess the perceptions of the postgraduate students about CSM for learning experimental pharmacology practical.

The present study is designed to evaluate the perception of postgraduate students toward animal simulation as a learning method for experimental pharmacology practical.

Materials and Methods

This was a cross-sectional, observational study conducted in the Department of Pharmacology, Seth GS Medical College and KEM Hospital, Parel in Mumbai, India. The study was initiated after obtaining Institutional Ethics Committee permission (EC/OA-116/2017) and written informed consent of the participants. In the pharmacology department, postgraduate students are posted in experimental laboratory for 2 months. Thirty study participants were 1st, 2nd, and 3rd year postgraduate students who had completed their routine 2 months' experimental pharmacology laboratory posting. For the first 15 days of experimental pharmacology posting, students read the resource material including CD or pen drive of CSM provided to them by teachers. Students are expected to conduct isolated tissue experiments using CSM. After 15 days, students conduct live isolated tissue experiments which include, i.e., rat colon, guinea pig ileum, rat fundus, and rat uterus. Those students who have used the animal simulation software and had completed at least five isolated tissue experiments were included in this study. At the end of the posting, students were given questionnaires, which was filled by them and submitted to the investigators after 25–30 min.

Study tools

The two important study tools in this research were the Animal simulator (Reed Elsevier India Pvt. Ltd.,) software and the study questionnaire.

Animal simulation software

The department of pharmacology and therapeutics has purchased animal simulation software from Elsevier. The pen drive provided by Elsevier consists of two softwares, namely Ex-Pharm and Xcology.

Ex-pharm

The package contains programs such as effects of drugs on the rabbit eye, effects of drugs on the frog heart, bioassay of histamine on the guinea pig ileum, effects of drugs on the frog esophagus, and effects of drugs on dog blood pressure and heart rate. These programs can simulate drug actions in various animal tissues. The user can conduct experiments and collect as well as analyze data. Each program can be run in two modes – a tutorial mode and an examination mode.[7]

X-cology

This software (developed in Multimedia Director, Flash and visual basic for Windows) displays complete video demonstrations of different procedures such as isolation and mounting of animal tissues followed by on-screen interactive interface to study the effects of various drugs on the isolated tissues.[8] The details on the experiments involving animal use are divided into different topics to facilitate their presentation and ease the navigation through the details. The content is classified into the following topics:

The experimental animals: This section includes biological names of the common experimental animals and their use in experimental pharmacology

The equipment: This section includes the common instruments used for the demonstration of animal experiments. Experimental techniques: This section includes the information on manual skills and routine procedures involved in the experimental pharmacology such as collection of blood samples, preparation of drug solutions, and routes of drug administration. The experimental section contains exhaustive details such as video demonstrations on isolation and mounting of different tissues from experimental animals, an interactive interface to study effects of different drugs on isolated tissues, procedures to carry out bioassays and experiments on whole animals related to screening and evaluation of drugs.

Questionnaire

A structured prevalidated questionnaire was used to evaluate the perception of students. The statements of the questionnaire were determined through multiple group discussions with 5–6 pharmacologists (who have used CSM in their teaching) and review of literature. The questionnaire consisted of 17 questions under the following domains:

Usage of CSM and live tissue experiments

This domain asked the students to provide information regarding types of tissues used and types of graphs/bioassay experiments performed using either CSM/live tissue experiments. Responses were in binary format, i.e., yes/no.

Logistics of conducting animal experiments

Statements regarding ethical issues, number of animals, time required, technical errors, and anxiety were included.

Advantages of CSM

Statements regarding repeatability of experiment, understanding (set up. Graph, calculations), retention of knowledge, visualization of effects of drugs were included in this domain.

Disadvantages of animal experiments in learning

Statements regarding the computer expertise, time, visibility of variation in responses were included in this domain.

Usefulness of CSM in practice

Statements pertaining to usefulness of CSM in UG/PG teaching were included in this domain.

Response for domain B, C, D, and E was in the form of three-point Likert Scale, i.e., agree, neutral, and disagree. At the end of the questionnaires, space was provided for the respondents to indicate their viewpoints and suggest any steps to improve this technique.

Questionnaire validation

Four faculty members in the pharmacology department and two medical education experts validated the questionnaire for the content. The validation method used was qualitative for the viewpoints on the relevancy or representativeness, clarity, and comprehensiveness of the items in the questionnaire by the experts. The inputs by the experts were incorporated into the questionnaire.

The questionnaire was validated quantitatively by Content Validity Ratio (CVR) by administering it to 10 subject experts.[910] Each expert assigned a scale to each question, i.e., essential, useful but not essential, and not essential. The CVR for each question was calculated by the Lawshe's Formula. According to this formula, a minimum CVR value of 0.62 is required for ten- panel members.

In our questionnaire, items having CVR value more than 0.62 were retained.

Statistical analysis

Graph Pad InStat, version 3.06 (GraphPad Software, Inc., San Diego, CA, USA) was used for statistical analysis. Descriptive analysis was used to assess the individual questions on student perception questionnaire. The responses were expressed as number/percentage of students stating yes/no, agreeing, disagreeing or are neutral for the particular statement. The number of students conducting experiments using CSM as compared to live animal experiments (LAE) for types of tissue and types of bioassay methods was compared using Chi-square test. P < 0.05 was considered statistically significant.

Results

Of the total 37 PG students approached (included students who were in the department and those who were serving 1-year postgraduate compulsory bond in nearby institutions), two students refused consent. Of the 35 students, 30 students returned completely filled questionnaires which were analyzed, data of which is presented in Tables 1 and 2.

Table 1

Number of students conducting experiments using live tissue and computer-simulated method (n=30)

Type of tissue	Live tissue taken (out of 30) (%)	CSM (out of 30) (%)	P
Rat colon	30 (100)	30 (100)	-
Guinea pig ileum	30 (100)	14 (46.67)	<0.001*
Rat uterus	19 (63.33)	14 (46.67)	0.29
Rat fundus	25 (83.33)	7 (23.33)	<0.001*
Frog rectus abdominis	-	3 (10)
Guinea pig tracheal chain	-	4 (13.33)
Rat phrenic nerve diaphragm	-	3 (10)
Frog esophagus	-	1 (3.33)
Rabbit jejunum	-	1 (3.33)
Rabbit aortic strips	-	1 (3.33)
Cat phrenic nerve diaphragm	-	1 (3.33)

*P<0.05 using Chi-square test. CSM=Computer-simulated method. P value or probaility value is the probaility for a given statistical model. A small p value (<0.05) indicates strong evidence against the null hypothesis

Table 2

Quantitative bioassay performed by study participants using live tissue/computer-simulated method (CSM)

Type of bioassay method	Live tissue taken (out of 30) (%)	CSM (out of 30) (%)	P
Graded DRC	24 (80)	29 (96.67)	0.10
Matching assay	30 (100)	18 (60)	<0.01*
Bracketing assay	25 (83.33)	14 (46.67)	0.006*
Interpolation assay	17 (56.67)	2 (6.67)	<0.001*
3-point assay	29 (96.67)	18 (60)	0.001*
4-point assay	9 (30)	2 (6.67)	0.041*
Determination of PA2	-	2 (6.67)
Determination of PD2	-	2 (6.67)
Cumulative DRC	-	2 (6.67)

*P<0.05 using Chi-square test. DRC=Dose response curve, CSM=Computer-simulated method

Domain A

Of the total 11 tissue experiments, four tissues were used for LAE, whereas with CSM, students could perform experiments for all tissues. Majority of students had performed experiments for four tissues, i.e., rat colon, guinea pig ileum, rat uterus, and rat fundus using both LAE and CSM. Among these tissues for which both LAE and CSM method was used, rat colon was performed by all students (n = 30) using both methods. Majority of students used LAE for guinea pig ileum (LAE: 30 vs. CSM: 14, P < 0.001), rat uterus (LAE: 19 vs. CSM: 14, P = 0.29), and rat fundus (LAE: 25 vs. CSM: 7, P < 0.001), as compared to CSM.

Of the remaining tissue experiments which were performed using only CSM, guinea pig tracheal chain was conducted by four students, frog rectus abdominis and rat phrenic nerve diaphragm by three students each and rest of the tissue experiments were performed by one student each [Table 1].

Out of the total nine bioassay methods enumerated in questionnaire, students had performed six methods using both LAE and CSM. Of these six bioassay methods, five bioassay methods were performed by majority of students using LAE compared to CSM, which included matching assay (LAE: 30 vs. CSM: 18, P < 0.001), bracketing assay (LAE: 25 vs. CSM: 14, P = 0.006), interpolation assay (LAE: 17 vs. CSM: 2, P < 0.001), three-point assay (LAE: 29 vs. CSM: 18, P = 0.001), and four-point assay (LAE: 9 vs. CSM: 2, P = 0.041). The graded dose-response curves were plotted by all students using both LAE as well as CSM methods. The other three bioassay methods, namely determination of PA2, determination of PD2, and cumulative DRC, were performed by two students each, using only CSM [Table 2].

Domain B, C, D, and E

When questioned about the logistics of conducting experiments, majority of the students opined that CSM overcomes ethical issues (agree: 29/30) and reduces number of animals used (agree: 26/30). Students felt that CSM reduces anxiety (agree: 23/30) as well as technical errors (agree: 23/30) related to LAE if used before it and is less time consuming (agree: 23/30) [Figure 1]. Most of the students did not agree that using CSM requires high level of computer expertise (disagree: 20/30), but they felt that variations in responses as seen in LAE cannot be mimicked by CSM (agree: 13/30) [Figure 2].

Figure 1

Domain B: Logistics of conducting experiments using either CSM or live animal experiments

Figure 2

Domain D: Perception regarding disadvantages of CSM compared to live animal experiments

Majority of the students agreed that CSM has advantages over LAE in terms of repeatability of experiments (agree: 26/30), clearer visualization (agree: 17/30), better understanding of isolated tissue set up (agree: 15/30), and easier calculations or graph plotting (agree: 19/30). However, the students disagreed with the statement that retention of knowledge is better with the use of CSM (disagree: 11/30) [Figure 3].

Figure 3

Domain C: Perception regarding advantage of CSM compared to live animal experiments

Most of the students opined that CSM can be used for difficult, lengthy experiments (agree: 19/30) and for undergraduate or postgraduate teaching (agree: 19/30). However, opinions regarding replacing LAE with CSM in PG teaching were divided (agree: 7, neutral: 12, and disagree: 12) [Figure 4].

Figure 4

Domain E: Perception regarding usefulness of CSM in practice

Discussion

Isolated tissue experiments form an important part of the curriculum for postgraduate students of pharmacology. Isolated tissue experiments have been used by pharmacologists from over 100 years for its versatility, simplicity, and reproducibility. The versatility of this method has allowed scientists to characterize receptors, which forms the basis for therapies of various diseases. However, this method requires sacrificing animals and has come under the scanner of animal rights activists.

The use of animals for experiments in medical education is complicated in India because of concerns raised by various regulatory bodies such as The CPCSEA, UGC, and Medical Council of India (MCI). The recent MCI directive asks medical colleges to use alternatives in undergraduate medical education, but there is no clarity from the authorities' end about the use of animals in postgraduate courses. Because of this, the use of animals across medical colleges in India has decreased for undergraduate teaching. However, as there are no clear-cut regulations or guidelines regarding the use of animals for postgraduate teaching, confusion persists. Different approaches are being implied to minimize the use of animals or to generate alternatives to experimental animals. However, the animal use has not been totally excluded. Some colleges have shifted to alternatives such as using computer simulations, while few institutions have still retained use of LAE in postgraduate syllabus.

Computer-simulated experiments have been used in pharmacological teaching and research for nearly two decades and proven to have several advantages over the traditional LAE, offering more accurate and consistent end results, along with flexibility as to when and where the experiments are conducted.

In our study, the perception of the postgraduate students in pharmacology regarding the use of CSM compared to LAE for isolated tissue experiments was evaluated. LAE were performed for only four tissues, i.e., rat colon, guinea pig ileum, rat uterus, and rat fundus, which were the same tissues chosen by maximum students for performing CSM procedures. This could be because examiners ask students to perform isolated tissue experiments using these four tissues most commonly during postgraduate examinations. Students may not prefer LAE for the other tissues which included frog, rabbit, guinea pig, and cat, as there are issues with the handling, killing, dissection, and availability of these animals. This shows that students prefer smaller animals such as rat and mice for live isolated tissue experiments. For these experiments involving frog, rabbit, guinea pig, cat, as well as for time-consuming experiments such as rat phrenic nerve-diaphragm, students used CSM method. However, the number of students venturing into these tissues using CSM was small.

Students preferred to perform assays which were less time consuming and less complicated (6 out of 11 assays). Again, majority of students preferred LAE for these six assays compared to CSM, thus indicating that these assays could be more important for their examinations. However, assays requiring expertise and more time (such as antagonistic assays and cumulative DRC) were performed using CSM by a small number of students.

In the present study, the analysis of feedback showed that majority of students perceived the importance of CSM for ethical aspect and reducing animal use. These findings, including the students' opinion regarding reduction of technique errors, time required and decreased anxiety related to LAE if used after CSM, are in line with the similar study by Sharma et al.[5] In this particular study, 95% of students had agreed to similar statements like those in our validated questionnaire. In the study by Kaur et al., 50% of faculty members from pharmacology department felt that CSM is time-consuming, in contrast to the opinion of postgraduate students in our study.[11]

In our study, there was clear agreement for the statements regarding the advantages of the CSM for understanding the concepts of experimental pharmacology and repeatability. These findings were compared to findings of the study by Sharma and Malhotra in which perception of both undergraduate students as well as faculty was obtained.[12] In this study, 99% of faculties felt that there is a better understanding with CSM whereas only 25% of the undergraduate students felt the same. A study by Babu et al. reported that 58.33% of undergraduate students perceived CSM is useful for better understanding of bioassay.[13] In our study, 63.33% of students agreed that CSM provides a better understanding of calculations and graphs and 50% agreed that CSM provides better understanding of the setup needed for isolated tissue experiments. The variation in findings could be because of difference in the level of experience, and difference in the expectations for the participants. The responses related to perception for repeatability (86.66%) and visualization of drug effects (56.66%) in our study are similar to those reported in the study by Babu et al. and Sharma et al., with majority of students agreeing. There was ambiguity in the response for the statement in our study questionnaire regarding increased retention of knowledge by CSM, with 36.33% showing disagreement and 40% having a neutral stand on the same. In a study by Atray et al., authors had compared CSM with conventional method for teaching pharmacology in undergraduate practicals, and found that majority of students had enhanced retaining capacity following CSM (97%) compared to conventional method (81%).[14] Similar enhanced retention capacity of CSM was perceived by undergraduate students in the studies by Sharma et al. and Babu et al.[513] Again, these findings point to the fact that postgraduate students prefer LAE for retention of knowledge and performing better in postgraduate practical examinations.

Few authors have reported that students and faculty felt need for computer expertise (Sharma et al.); however, postgraduate students in our study did not feel the same. However, the most important disadvantage emphasized by all studies is the variation in responses which is difficult to visualize by CSM, as there are prefixed doses involved and there is no interaction with the live tissue.[512] In our study, there was ambiguity regarding responses for this statement, with 50% showing disagreement and 40% having neutral responses. This indicates that many postgraduate students find it difficult to decide which method is better for visualizing variations in response. The reason for this could be the short period of CSM exposure in our institute, as only 15 days are given during the posting for postgraduate students to understand and perform experiments on CSM software before proceeding to LAE.

There was a clear cut agreement (63%) for using CSM for lengthy/difficult experiments and using CSM for undergraduate or postgraduate pharmacology practicals. However, when asked if CSM should replace LAE for isolated tissue experiments, the opinions were divided with approximately 33% agreeing and an equal number of participants disagreeing on the same. Thus, there was difficulty for the students to choose clearly between CSM and LAE.

In contrast to most of the studies, where undergraduate students gave preference to CSM over LAE for pharmacology practical, the opinion of postgraduate students was inclined toward the use of both LAE with CSM in our study. In a study conducted by Shehnaz et al. where the perception of postgraduate faculty toward LAE was obtained, 60% of faculty were in favor of using LAE while 40% wanted to use alternatives for LAE.[15] Similarly, in a study conducted by Saache et al., 60% of postgraduate students and 83.3% of teachers agreed with the use of animal experiments in current pharmacology curriculum, especially procedures like bioassay.[16] However, among those in favor of LAE for pharmacology, 60% of teachers were not in favor of animal experiments for undergraduates.

Limitations of the study

As there was no published literature regarding the perception of postgraduate students in pharmacology toward CSM and LAE, we compared our finding with studies involving undergraduate students or those involving pharmacology faculty. The study can be conducted in larger sample size to obtain a better picture of the opinions.

Conclusion

CSM should be integrated alongside LAE to complement, reinforce, and enhance learning about bioassay experiments. Feedback from students must be examined with great deliberation. Regulatory bodies such as the MCI, CPCSEA, UGC, and ICMR should finalize uniform guidelines for conducting animal experiments along with virtual experiments in postgraduate practical pharmacology.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.